Animal collagen from non-animal sources

Abstract

The present invention relates to a non-animal organism expressing a collagen that naturally occurs extracellularly in one or more animal species. Furthermore, the invention refers to methods for preparing such collagen and for preparing a comestible nutrient product comprising such collagen obtained from a method of the present invention.

Description

[0001] Animal collagen from non-animal sources

[0002] The present invention relates to a non-animal organism expressing a collagen. A non-animal organism expressing a collagen that naturally occurs extracellularly in one or more animal species. Furthermore, the invention refers to methods for preparing such collagen and for preparing a comestible nutrient product comprising such collagen. Furthermore, the invention relates to a comestible nutrient product comprising such collagen obtained from a method of the present invention.

[0003] There is a considerable global demand for animal protein in the fields of nutrition as well as pharmaceutical uses. However, there are increasing ethical and economic constraints and concerns when using such proteins obtained from slaughtered animals. Furthermore, undesirable material such as xenobiotics, hormones and precursors thereof and high caloric ingredients are inevitably present in animal- derived polypeptides, such as collagen, depending on the growth conditions of the animals.

[0004] Therefore, there is a considerable interest for obtaining polypeptides of animal origin (animal-like polypeptides such as collagen) obtained from other sources without the need of slaughtering an animal.

[0005] It was surprisingly found that a number of collagens that naturally occur extracellularly in one or more animal species can efficiently be obtained from expression in non-animal organism of various species of various living being kingdoms such as such as plants, fungi and bacteria. This is experimentally evidenced as shown below.

[0006] An aspect of the present invention relates to a non-animal organism expressing a collagen that naturally occurs extracellularly in one or more animal species. The term non-animal organism does not include humans.

[0007] In other words, the present invention relates to a non-animal organism expressing a collagen that originates from an animal species in the animal’s extracellular space. The term “non-animal organism” may be understood in the broadest sense as an organism that is not of animal origin. Thus, the term “non-animal organism” may be understood in the broadest sense as a living being of any biological kingdom, excluding an animal, such as, e.g., an organ or tissue of one or more plants, one or more fungi, one or more protista, one or more eubacteria, and / or one or more archaebacterial. The non-animal organism may be a monocellular organism or may be a multicellular organism, which optionally has different cell types and / or tissues. In one embodiment, the non-animal organism is a protist. In one embodiment, the non-animal organism is a eukaryote. Using a eukaryote may have the advantage that posttranslational modifications may be included.

[0008] As used herein, the term “animal” may be understood in the broadest sense as commonly understood in the art. Typically, an animal is a multicellular, eukaryotic organism in the biological kingdom Animalia, typically consuming organic material and breathing oxygen, and having myocytes as typical cell types. Typically, animal cells do not have a cell wall, neither of cellulose, hemicellulose nor of chitin.

[0009] The terms “organism”, “living being”, “creature”, and “living creature” may be understood in the broadest sense as a multicellular organism or a single cellular organism. In a preferred embodiment, the non-animal organism is selected from the group consisting of a plant, a fungus, a protist, a bacterium, and an archaebacterium. In a preferred embodiment, the non-animal organism is selected from the group consisting of a plant, a fungus, and a bacterium.

[0010] In a preferred embodiment, the non-animal organism is selected from the group consisting of:

[0011] (A) a plant, in particular a cultivated plant, in particular tobacco or soy;

[0012] (B) a fungus, preferably a yeast cell, in particular Pichia pastoris', or

[0013] (C) a bacterium, in particular an Escherichia coli bacterium.

[0014] In a preferred embodiment, the non-animal organism is a plant. In a preferred embodiment, the non-animal organism is a fungi, in particular yeast. In a preferred embodiment, the non-animal organism is a bacterium, in particular Escherichia coli.

[0015] As used herein, a polypeptide, in particular collagen, that naturally occurs extracellularly in one or more animal species may be understood in the broadest sense as any polypeptide, in particular collagen, that is physiologically located in the extracellular space of at least one animal species. Such extracellular space may be any location outside the cell lumen and cell membrane. An extracellular space may be an extracellular matrix that may be solid.

[0016] In the context of the present invention, the terms “polypeptide” and “protein”, in particular when used in the context of collagen in the present invention, may be understood interchangeably in the broadest sense as a compound mainly composed of natural amino acid moieties consecutively conjugated with one another via amide bonds. It will be understood that a protein in the sense of the present invention may or may not be subjected to one or more posttranslational modifications and / or be conjugated with one or more non-amino acid moiety / moieties. The termini of the protein may optionally be capped by any means known in the art, such as, e.g., amidation, acetylation, methylation, and / or acylation. Posttranslational modifications are well-known in the art and may be, but may not be limited to, lipidation, phosphorylation, sulfatation, glycosylation, truncation, oxidation, reduction, decarboxylation, acetylation, amidation, deamidation, disulfide bond formation, hydroxylation, amino acid addition, cofactor addition (e.g., biotinylation, heme addition, eicosanoid addition, steroid addition) and complexation of metal ions, non-metal ions, peptides or small molecules and addition of ironsulphide clusters. Moreover, optionally, co-factors, in particular cyclic guanidinium monophosphate (cGMP), but optionally also such as, e.g., ATP, ADP, NAD+, NADH+H+, NADP+, NADPH+H+, metal ions, anions, lipids, etc. may be bound to the protein, irrespective on the biological influence of these co-factors. It will be understood that such polypeptide, such as collagen, may also bear one or more non-natural amino acid moiety / moieties and / or one or more posttranscriptional modifications. The one or more optional posttranslational modifications typically depend on the non-animal organism in which the polypeptide, such as collagen, of interest is expressed.

[0017] The expression of the polypeptide, such as collagen, may be permanent or may be inducible. This may be adjusted by the choice of a promoter and / or enhancer. In a case where the promoter and / or enhancer is permanently active, the polypeptide, such as collagen, may be permanently expressed. When the promoter and / or enhancer is controllable by a certain component, the non-animal organism may be grown without expression of the protein and triggering expression whenever it is desired.

[0018] The collagen may be expressed in any form. For instance, it may be provided in a gel-like or liquid composition, or may be a secret (also: secretion), preferably comprised in a secret, in particular a secret from a gland. As used herein, the terms “secret” and “secretion” may be understood interchangeably.

[0019] In a preferred embodiment, the collagen is selected from the list consisting of extracellular matrix of:

[0020] (a) skin;

[0021] (b) tendon;

[0022] (c) bone,

[0023] (d) teeth,

[0024] (e) cartilage,

[0025] (f) ligaments,

[0026] (g) blood vessels and

[0027] (h) a combination of two or more thereof.

[0028] The collagen that is expressed by the non-animal cell may be of any origin. In other words, it may be any animal collagen. The term "... collagen” may be understood in the broadest sense being naturally of this origin or a collagen of high homology (or identity) of at least 80%, of at least 85%, of at least 90%, of at least 95%, of at least 98%, of at least 99%, or 100% being naturally of this origin. This may include native and mutated collagen types. In other words, the respective collagen or a highly homologous collagen thereof is typically found and / or expressed in the respective species. Typically, the collagen or a precursor thereof is genetically encoded in the respective animal organism. Optionally, it may be based on a spliced genetic sequence and / or may be subjected to posttranslational splicing.

[0029] Collagens are well-known polypeptides, which typically bear a considerable content of proline. Collagen may be any type of collagen such as, e.g., COL1 (e.g., COL1 A1 or COL1A2) or COL2. Collagen may form hydroxylated heterodimeric helices. Lysine residues may be hydroxylated (e.g., by lysyl hydroxylase (LH)). In addition, proline may be hydroxylated (e.g., by prolyl-4-hydroxylase (P4H)).

[0030] In a preferred embodiment, the collagen that is expressed by the non-animal cell is a vertebrate collagen. In a preferred embodiment, the collagen that is expressed by the non-animal cell is a mammal collagen, including a human or non-human collagen, a bird collagen, a reptile collagen, or a fish collagen. In a preferred embodiment, the collagen that is expressed by the non-animal cell is a mammal collagen, including a human or non-human collagen, or a bird collagen. In a preferred embodiment, the collagen that is expressed by the non-animal cell is a human, a bovine, a porcine, a sheep, a goat, a horse, a donkey, or a chicken collagen.

[0031] Alternatively, it may also be an invertebrate collagen such as an arthropod collagen such as, e.g., an insect or crustacean collagen.

[0032] In a preferred embodiment, the collagen that is expressed by the non-animal cell has at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% homology (or identity) of an extracellular matrix collagen such as collagen, more preferably wherein the extracellular matrix collagen is selected from mammal extracellular matrix collagen and bird extracellular matrix collagen.

[0033] In a preferred embodiment, the collagen is of comestible grade. In a preferred embodiment, the collagen is of pharmaceutical grade.

[0034] In a preferred embodiment, the collagen that is expressed by the non-animal cell has at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% homology (or identity) of bovine collagen (COL1A1 , collagen alpha 1 (1)) (>sp|P02453|CO1 A1_BOVIN Collagen alpha-1 (I) chain OS=Bos taurus OX=9913 GN=COL1A1 PE=1 SV=3) of SEQ ID NO: 1 :

[0035] MFSFVDLRLLLLLAATALLTHGQEEGQEEGQEEDIPPVTCVQNGLRYHDRDVWK PVPCQICVCDNGNVLCDDVICDELKDCPNAKVPTDECCPVCPEGQESPTDQETT GVEGPKGDTGPRGPRGPAGPPGRDGIPGQPGLPGPPGPPGPPGPPGLGGNFA PQLSYGYDEKSTGISVPGPMGPSGPRGLPGPPGAPGPQGFQGPPGEPGEPGA SGPMGPRGPPGPPGKNGDDGEAGKPGRPGERGPPGPQGARGLPGTAGLPGM KGHRGFSGLDGAKGDAGPAGPKGEPGSPGENGAPGQMGPRGLPGERGRPGA PGPAGARGNDGATGAAGPPGPTGPAGPPGFPGAVGAKGEGGPQGPRGSEGP QGVRGEPGPPGPAGAAGPAGNPGADGQPGAKGANGAPGIAGAPGFPGARGP SGPQGPSGPPGPKGNSGEPGAPGSKGDTGAKGEPGPTGIQGPPGPAGEEGKR GARGEPGPAGLPGPPGERGGPGSRGFPGADGVAGPKGPAGERGAPGPAGPK GSPGEAGRPGEAGLPGAKGLTGSPGSPGPDGKTGPPGPAGQDGRPGPPGPP GARGQAGVMGFPGPKGAAGEPGKAGERGVPGPPGAVGPAGKDGEAGAQGPP GPAGPAGERGEQGPAGSPGFQGLPGPAGPPGEAGKPGEQGVPGDLGAPGPS GARGERGFPGERGVQGPPGPAGPRGANGAPGNDGAKGDAGAPGAPGSQGAP GLQGMPGERGAAGLPGPKGDRGDAGPKGADGAPGKDGVRGLTGPIGPPGPAG APGDKGEAGPSGPAGPTGARGAPGDRGEPGPPGPAGFAGPPGADGQPGAKG EPGDAGAKGDAGPPGPAGPAGPPGPIGNVGAPGPKGARGSAGPPGATGFPGA AGRVGPPGPSGNAGPPGPPGPAGKEGSKGPRGETGPAGRPGEVGPPGPPGP

[0036] AGEKGAPGADGPAGAPGTPGPQGIAGQRGWGLPGQRGERGFPGLPGPSGEP GKQGPSGASGERGPPGPMGPPGLAGPPGESGREGAPGAEGSPGRDGSPGAK GDRGETGPAGPPGAPGAPGAPGPVGPAGKSGDRGETGPAGPAGPIGPVGARG PAGPQGPRGDKGETGEQGDRGIKGHRGFSGLQGPPGPPGSPGEQGPSGASG

[0037] PAGPRGPPGSAGSPGKDGLNGLPGPIGPPGPRGRTGDAGPAGPPGPPGPPGP PGPPSGGYDLSFLPQPPQEKAHDGGRYYRADDANVVRDRDLEVDTTLKSLSQQ IENIRSPEGSRKNPARTCRDLKMCHSDWKSGEYWIDPNQGCNLDAIKVFCNMET GETCVYPTQPSVAQKNWYISKNPKEKRHVWYGESMTGGFQFEYGGQGSDPAD

[0038] VAIQLTFLRLMSTEASQNITYHCKNSVAYMDQQTGNLKKALLLQGSNEIEIRAEGN SRFTYSVTYDGCTSHTGAWGKTVIEYKTTKTSRLPIIDVAPLDVGAPDQEFGFDV GPACFL

[0039] The sequence without propeptides may be of SEQ ID NO: 2:

[0040] QLSYGYDEKSTGISVPGPMGPSGPRGLPGPPGAPGPQGFQGPPGEPGEPGAS GPMGPRGPPGPPGKNGDDGEAGKPGRPGERGPPGPQGARGLPGTAGLPGMK GHRGFSGLDGAKGDAGPAGPKGEPGSPGENGAPGQMGPRGLPGERGRPGAP GPAGARGNDGATGAAGPPGPTGPAGPPGFPGAVGAKGEGGPQGPRGSEGPQ

[0041] GVRGEPGPPGPAGAAGPAGNPGADGQPGAKGANGAPGIAGAPGFPGARGPS GPQGPSGPPGPKGNSGEPGAPGSKGDTGAKGEPGPTGIQGPPGPAGEEGKRG ARGEPGPAGLPGPPGERGGPGSRGFPGADGVAGPKGPAGERGAPGPAGPKG SPGEAGRPGEAGLPGAKGLTGSPGSPGPDGKTGPPGPAGQDGRPGPPGPPGA

[0042] RGQAGVMGFPGPKGAAGEPGKAGERGVPGPPGAVGPAGKDGEAGAQGPPGP AGPAGERGEQGPAGSPGFQGLPGPAGPPGEAGKPGEQGVPGDLGAPGPSGA RGERGFPGERGVQGPPGPAGPRGANGAPGNDGAKGDAGAPGAPGSQGAPGL QGMPGERGAAGLPGPKGDRGDAGPKGADGAPGKDGVRGLTGPIGPPGPAGAP

[0043] GDKGEAGPSGPAGPTGARGAPGDRGEPGPPGPAGFAGPPGADGQPGAKGEP GDAGAKGDAGPPGPAGPAGPPGPIGNVGAPGPKGARGSAGPPGATGFPGAAG RVGPPGPSGNAGPPGPPGPAGKEGSKGPRGETGPAGRPGEVGPPGPPGPAG EKGAPGADGPAGAPGTPGPQGIAGQRGWGLPGQRGERGFPGLPGPSGEPGK

[0044] QGPSGASGERGPPGPMGPPGLAGPPGESGREGAPGAEGSPGRDGSPGAKGD RGETGPAGPPGAPGAPGAPGPVGPAGKSGDRGETGPAGPAGPIGPVGARGPA GPQGPRGDKGETGEQGDRGIKGHRGFSGLQGPPGPPGSPGEQGPSGASGPA GPRGPPGSAGSPGKDGLNGLPGPIGPPGPRGRTGDAGPAGPPGPPGPPGPPG

[0045] PPSGGYDLSFLPQPPQEKAHDGGRYYRA In a preferred embodiment, the collagen that is expressed by the non-animal cell may be based on a DNA sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% homology (or identity) of a sequence for the expression in E. coli within pET-21 (+) from Twist Bioscience without propeptides (RBS with ATG from pET 151 ) of SEQ ID NO: 3:

[0046] TAAGAAGGAGATATACATATGCATCACCATCACCATCACGAAAACCTGTATTT TCAGGGC

[0047] CAACTGTCATATGGCTATGACGAGAAAAGCACCGGTATATCTGTCCCGGGGC CAATGGGCCCATCAGGACCACGCGGTTTGCCAGGCCCACCGGGTGCCCCG GGTCCTCAAGGATTCCAAGGACCACCCGGTGAACCAGGTGAACCGGGTGCT TCTGGCCCGATGGGTCCTAGAGGTCCGCCGGGGCCCCCAGGCAAGAATGGA GATGATGGTGAAGCGGGTAAACCAGGCAGACCCGGTGAGAGAGGTCCCCCG GGACCACAGGGCGCTCGCGGACTTCCAGGTACAGCGGGATTGCCTGGAATG AAAGGGCATCGTGGCTTTTCTGGCTTGGATGGAGCAAAAGGCGATGCTGGG CCAGCAGGTCCAAAGGGTGAACCAGGAAGTCCAGGAGAAAATGGTGCTCCT GGTCAAATGGGACCCCGTGGGTTACCAGGTGAGCGGGGACGTCCAGGAGCT CCCGGCCCCGCAGGTGCTCGCGGTAACGACGGTGCCACCGGTGCGGCTGG ACCACCGGGTCCTACAGGGCCGGCAGGGCCCCCTGGATTTCCTGGTGCAGT AGGTGCGAAGGGAGAAGGTGGACCTCAAGGTCCCCGAGGATCGGAAGGCC CCCAAGGAGTCCGCGGTGAGCCAGGTCCGCCGGGTCCTGCTGGTGCAGCA

[0048] GGGCCGGCTGGTAATCCAGGTGCGGATGGACAGCCTGGTGCCAAAGGTGCA AATGGTGCTCCGGGGATTGCCGGCGCCCCAGGATTTCCCGGTGCTAGAGGT CCATCTGGCCCGCAAGGTCCTTCTGGACCGCCGGGCCCTAAAGGCAACAGC GGTGAACCAGGGGCTCCCGGAAGCAAAGGAGATACCGGAGCCAAGGGAGA ACCGGGCCCGACCGGGATTCAGGGCCCACCGGGTCCAGCAGGCGAAGAAG

[0049] GTAAACGCGGTGCACGTGGGGAACCAGGTCCTGCAGGCTTACCGGGTCCCC CTGGTGAGAGAGGCGGTCCGGGTTCACGAGGCTTTCCAGGAGCGGATGGAG TGGCAGGCCCTAAAGGGCCGGCGGGAGAGCGGGGCGCCCCTGGCCCCGC AGGTCCAAAAGGGTCTCCAGGTGAAGCTGGGCGTCCGGGTGAAGCAGGATT GCCTGGTGCAAAGGGGTTAACTGGCAGTCCTGGTAGTCCGGGACCGGATGG

[0050] TAAAACTGGTCCGCCGGGCCCGGCAGGTCAAGATGGACGTCCAGGACCCCC GGGACCCCCTGGAGCTCGAGGTCAAGCTGGAGTAATGGGCTTTCCGGGTCC GAAGGGAGCAGCGGGCGAACCCGGAAAGGCAGGCGAACGGGGAGTTCCAG GTCCACCCGGTGCAGTAGGCCCAGCAGGGAAAGATGGCGAGGCTGGTGCT CAAGGGCCACCTGGTCCAGCAGGCCCGGCTGGCGAACGTGGAGAACAAGG

[0051] ACCGGCGGGAAGCCCTGGATTTCAGGGATTGCCTGGCCCAGCGGGACCCCC TGGAGAGGCGGGTAAACCAGGAGAACAAGGTGTACCAGGAGATCTTGGTGC

[0052] ACCTGGTCCTAGCGGAGCTCGTGGTGAAAGAGGTTTTCCAGGTGAGCGCGG

[0053] TGTTCAAGGTCCCCCGGGGCCAGCAGGACCTCGTGGAGCAAATGGTGCGCC

[0054] AGGTAACGATGGCGCAAAAGGTGATGCCGGTGCACCAGGTGCACCAGGGTC

[0055] ACAGGGTGCTCCCGGGTTACAAGGAATGCCAGGTGAGAGAGGTGCCGCAGG

[0056] GTTACCTGGGCCGAAGGGCGATAGAGGCGACGCTGGACCAAAAGGTGCCGA

[0057] TGGAGCTCCGGGTAAGGATGGAGTCAGAGGTTTAACAGGACCAATAGGACC

[0058] CCCAGGTCCTGCTGGTGCCCCTGGCGATAAAGGTGAGGCAGGTCCTAGTGG

[0059] TCCAGCCGGTCCCACAGGTGCACGCGGAGCTCCAGGTGATCGCGGCGAACC

[0060] GGGACCACCGGGGCCTGCTGGGTTTGCGGGTCCACCTGGTGCCGATGGAC

[0061] AACCTGGTGCGAAGGGCGAACCCGGTGATGCAGGCGCGAAGGGCGATGCA

[0062] GGACCACCAGGACCAGCTGGCCCGGCAGGTCCGCCGGGTCCTATCGGAAA

[0063] CGTCGGTGCGCCCGGGCCTAAAGGTGCTAGAGGGTCTGCAGGCCCGCCAG

[0064] GTGCTACTGGTTTCCCTGGTGCAGCTGGTCGTGTAGGGCCACCAGGCCCTT

[0065] CAGGAAATGCTGGTCCGCCTGGACCACCTGGGCCAGCAGGAAAAGAGGGGA

[0066] GTAAAGGACCACGTGGTGAGACAGGTCCGGCTGGCCGGCCTGGTGAAGTCG

[0067] GTCCACCTGGTCCGCCCGGACCCGCTGGAGAGAAAGGTGCACCGGGTGCA

[0068] GACGGTCCGGCTGGAGCGCCTGGAACTCCAGGCCCTCAAGGAATCGCCGG

[0069] CCAAAGAGGCGTAGTTGGTTTACCGGGTCAGCGCGGTGAACGTGGTTTTCCT

[0070] GGACTTCCTGGTCCTTCCGGTGAACCGGGTAAGCAAGGACCGTCTGGAGCA

[0071] AGCGGGGAACGTGGACCACCGGGCCCGATGGGTCCACCTGGGTTAGCTGG

[0072] TCCACCGGGCGAGTCTGGTAGAGAAGGCGCACCCGGGGCAGAAGGTTCAC

[0073] CAGGACGAGACGGCTCACCAGGGGCCAAAGGTGATAGAGGCGAAACAGGA

[0074] CCAGCGGGCCCGCCGGGAGCACCTGGTGCGCCGGGCGCTCCAGGTCCAGT

[0075] TGGACCGGCTGGAAAGTCAGGCGATAGAGGGGAAACGGGCCCGGCGGGAC

[0076] CAGCCGGCCCTATTGGACCAGTAGGAGCTCGTGGGCCGGCGGGGCCACAA

[0077] GGCCCTCGTGGAGATAAAGGTGAGACCGGTGAACAGGGAGATCGTGGGATT

[0078] AAGGGTCATCGAGGATTTTCAGGGCTTCAAGGACCGCCTGGTCCACCGGGC

[0079] TCGCCAGGTGAGCAGGGTCCCTCTGGTGCTTCTGGACCAGCTGGTCCACGT

[0080] GGCCCGCCAGGCAGTGCAGGCTCGCCAGGTAAAGATGGTCTTAACGGGTTA

[0081] CCTGGCCCAATTGGGCCACCAGGTCCTCGTGGTCGTACAGGCGATGCAGGT

[0082] CCAGCAGGTCCGCCTGGACCGCCAGGACCGCCTGGTCCCCCTGGTCCCCCA

[0083] TCAGGCGGATATGACCTTTCATTTCTCCCTCAGCCACCGCAAGAAAAGGCTC

[0084] ATGATGGTGGACGTTATTATCGCGCTTAA

[0085] In a preferred embodiment, the collagen that is expressed by the non-animal cell may be based on a DNA sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% homology (or identity) of codon- optimized sequence without tags or propeptides of SEQ ID NO: 4:

[0086] CAGCTGTCTTACGGGTATGATGAAAAAAGCACCGGCATTTCAGTTCCGGGCC

[0087] CGATGGGCCCGAGCGGTCCCCGTGGCCTGCCGGGCCCGCCGGGCGCGCC

[0088] GGGTCCGCAGGGCTTCCAAGGCCCGCCGGGCGAACCGGGTGAACCGGGCG

[0089] CCAGCGGCCCGATGGGCCCACGTGGCCCGCCGGGCCCGCCGGGCAAAAAT

[0090] GGCGATGATGGAGAAGCCGGCAAACCGGGCCGTCCGGGTGAACGTGGCCC

[0091] GCCGGGTCCGCAGGGCGCGCGCGGCCTGCCGGGTACCGCCGGCCTGCCG

[0092] GGCATGAAAGGTCATCGCGGCTTCAGCGGCCTGGATGGCGCCAAAGGTGAT

[0093] GCGGGTCCGGCGGGCCCGAAAGGCGAACCGGGCAGTCCGGGCGAAAATGG

[0094] CGCCCCGGGCCAGATGGGTCCGCGCGGTCTGCCGGGTGAACGTGGCCGCC

[0095] CGGGCGCGCCGGGTCCGGCAGGCGCGCGCGGCAACGATGGCGCGACCGG

[0096] CGCCGCCGGCCCGCCGGGCCCGACCGGCCCGGCGGGCCCGCCGGGATTT

[0097] CCGGGTGCTGTGGGCGCGAAAGGCGAAGGTGGCCCGCAGGGCCCGCGTGG

[0098] TAGCGAAGGTCCGCAGGGCGTTCGCGGCGAACCGGGCCCGCCGGGCCCGG

[0099] CAGGCGCGGCCGGCCCGGCGGGCAATCCGGGTGCCGATGGCCAGCCGGG

[0100] GGCCAAAGGCGCCAATGGTGCGCCGGGCATCGCGGGTGCACCGGGTTTTC

[0101] CGGGCGCGCGCGGCCCGAGCGGCCCGCAGGGCCCGAGCGGTCCGCCGGG

[0102] CCCGAAGGGCAACAGCGGCGAACCGGGCGCCCCGGGCAGCAAAGGCGATA

[0103] CCGGCGCGAAAGGCGAACCTGGCCCGACCGGCATTCAGGGCCCGCCGGGC

[0104] CCGGCAGGCGAAGAAGGCAAACGTGGCGCCCGCGGCGAACCGGGCCCGG

[0105] CGGGTCTGCCGGGCCCGCCGGGCGAACGTGGCGGCCCGGGCAGTCGCGG

[0106] CTTTCCGGGCGCCGATGGCGTGGCAGGCCCGAAAGGTCCGGCGGGCGAAC

[0107] GTGGCGCCCCGGGCCCGGCGGGCCCGAAAGGCAGCCCGGGGGAAGCGGG

[0108] CCGTCCGGGTGAAGCCGGCCTGCCGGGCGCGAAAGGCCTGACCGGCTCGC

[0109] CGGGCAGCCCGGGCCCGGATGGCAAAACCGGCCCGCCGGGCCCGGCAGG

[0110] CCAGGACGGCCGTCCGGGCCCGCCGGGCCCGCCGGGCGCCCGCGGTCAG

[0111] GCCGGCGTCATGGGCTTTCCGGGTCCGAAAGGCGCGGCGGGCGAACCGGG

[0112] TAAAGCGGGCGAACGTGGCGTGCCGGGCCCGCCGGGCGCCGTTGGCCCGG

[0113] CAGGCAAAGATGGCGAAGCGGGCGCCCAGGGTCCGCCGGGCCCGGCGGG

[0114] CCCGGCCGGCGAACGCGGCGAACAGGGCCCGGCGGGCAGCCCGGGTTTTC

[0115] AGGGCCTGCCGGGCCCGGCAGGCCCGCCGGGCGAAGCCGGTAAACCGGG

[0116] CGAACAGGGCGTGCCTGGCGATCTGGGCGCGCCGGGCCCGTCGGGAGCCC

[0117] GTGGTGAACGCGGCTTCCCGGGTGAACGTGGTGTGCAGGGCCCGCCGGGC

[0118] CCTGCCGGCCCGCGTGGCGCGAACGGCGCGCCGGGCAACGATGGCGCGAA

[0119] AGGCGACGCCGGCGCCCCGGGTGCGCCGGGCAGCCAGGGCGCGCCGGGC

[0120] CTGCAGGGTATGCCGGGCGAACGTGGCGCGGCGGGTCTGCCGGGCCCGAA AGGCGATCGCGGTGATGCTGGCCCGAAAGGCGCGGATGGCGCGCCGGGCA AGGATGGTGTGCGTGGCCTGACCGGCCCGATTGGCCCGCCGGGCCCGGCA GGCGCCCCGGGCGATAAAGGAGAAGCGGGCCCGAGCGGCCCGGCCGGCC CGACCGGTGCCCGTGGTGCGCCGGGCGATCGTGGCGAACCGGGTCCGCCG GGTCCGGCCGGCTTCGCGGGCCCGCCGGGCGCGGATGGCCAGCCGGGCG CGAAAGGCGAACCTGGCGATGCCGGTGCAAAGGGAGACGCGGGCCCTCCG GGCCCGGCGGGCCCGGCCGGACCACCGGGCCCGATTGGCAATGTTGGCGC GCCGGGCCCGAAAGGCGCACGTGGTAGCGCGGGCCCTCCGGGCGCCACCG GCTTTCCGGGCGCGGCCGGCCGTGTGGGCCCGCCGGGCCCGAGCGGCAAC GCGGGACCGCCGGGCCCGCCGGGCCCGGCCGGCAAAGAAGGCAGCAAAG GCCCGCGTGGCGAAACTGGCCCGGCCGGCCGTCCGGGCGAAGTTGGCCCG CCGGGCCCGCCGGGCCCGGCCGGTGAAAAAGGTGCCCCGGGTGCCGACG GCCCGGCGGGCGCACCGGGTACCCCGGGTCCGCAGGGTATTGCGGGCCAG CGCGGCGTTGTTGGTCTGCCGGGTCAACGTGGCGAACGCGGTTTCCCGGGC CTGCCGGGTCCGAGCGGCGAACCGGGTAAACAGGGCCCCAGCGGCGCCTC GGGCGAACGCGGTCCGCCGGGCCCGATGGGCCCGCCGGGCCTGGCGGGC CCGCCGGGCGAAAGTGGCCGCGAAGGCGCGCCGGGTGCAGAAGGCAGTCC GGGCCGTGATGGTTCACCGGGTGCGAAAGGTGATCGCGGCGAGACAGGTC CGGCCGGCCCGCCGGGCGCGCCGGGCGCCCCGGGCGCACCGGGTCCGGT CGGTCCGGCGGGCAAATCAGGAGATCGCGGCGAAACCGGTCCGGCGGGTC CGGCGGGCCCGATTGGTCCGGTGGGCGCGCGTGGCCCGGCAGGCCCGCA GGGTCCGCGCGGTGATAAAGGCGAAACCGGCGAGCAGGGCGATCGCGGTA TTAAAGGCCATCGTGGTTTCAGCGGTCTGCAAGGCCCGCCGGGCCCGCCGG GCTCGCCGGGCGAACAGGGCCCGTCGGGTGCGTCCGGCCCTGCCGGTCCG CGCGGCCCGCCGGGCAGCGCCGGCAGCCCGGGCAAAGATGGTTTAAACGG CCTGCCGGGCCCGATTGGCCCGCCGGGCCCGCGCGGCCGCACCGGCGATG CCGGCCCGGCAGGCCCGCCGGGCCCGCCGGGCCCGCCGGGTCCGCCGGG CCCGCCGAGCGGCGGCTATGATCTGAGCTTTCTGCCGCAGCCGCCGCAGGA AAAAGCGCATGATGGTGGCCGCTACTATCGAGCGTAA

[0121] In a preferred embodiment, the collagen that is expressed by the non-animal cell may be based on a DNA sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% homology (or identity) of codon- optimized sequence for the expression in P. pastoris rom VB with His-tag and TEV of SEQ ID NO: 5: ATGAGATTTCCTTCAATTTTTACTGCTGTTTTATTCGCAGCATCCTCCGCATTA

[0122] GCTGCTCCAGTCAACACTACAACAGAAGATGAAACGGCACAAATTCCGGCTG

[0123] AAGCTGTCATCGGTTACTCAGATTTAGAAGGGGATTTCGATGTTGCTGTTTTG

[0124] CCATTTTCCAACAGCACAAATAACGGGTTATTGTTTATAAATACTACTATTGCC

[0125] AGCATTGCTGCTAAAGAAGAAGGGGTATCTCTCGAGAAAAGAGAGGCTGAAG

[0126] CTCATCACCATCACCATCACGAAAACCTGTATTTTCAGGGCCAATTGTCTTAT

[0127] GGTTACGATGAAAAGTCCACTGGTATTTCTGTCCCTGGACCAATGGGTCCAT

[0128] CAGGTCCTAGAGGTTTGCCAGGTCCTCCAGGTGCTCCAGGTCCACAAGGTTT

[0129] TCAAGGTCCACCAGGTGAACCAGGAGAACCCGGTGCTTCTGGTCCAATGGG

[0130] TCCAAGAGGTCCACCTGGACCTCCTGGTAAGAACGGTGATGATGGTGAGGCT

[0131] GGTAAGCCAGGAAGACCTGGAGAAAGAGGACCACCTGGTCCACAAGGAGCT

[0132] AGAGGTTTACCCGGTACAGCTGGTTTGCCTGGTATGAAGGGTCATAGAGGTT

[0133] TCTCCGGTTTGGATGGAGCTAAGGGTGATGCTGGACCTGCTGGTCCTAAGG

[0134] GAGAGCCAGGATCTCCAGGTGAGAACGGAGCTCCTGGACAAATGGGACCAA

[0135] GAGGTCTGCCTGGTGAAAGAGGTAGACCAGGTGCTCCTGGTCCTGCCGGAG

[0136] CCAGAGGTAATGATGGTGCTACTGGTGCTGCTGGTCCTCCAGGTCCAACTGG

[0137] TCCTGCTGGTCCTCCAGGTTTTCCAGGTGCTGTTGGAGCCAAAGGTGAAGGT

[0138] GGCCCACAAGGTCCTAGAGGATCTGAAGGTCCACAGGGTGTTAGAGGTGAA

[0139] CCAGGTCCACCAGGTCCAGCTGGAGCTGCTGGTCCTGCTGGTAACCCAGGT

[0140] GCAGATGGTCAACCAGGTGCTAAGGGTGCCAACGGTGCTCCAGGCATTGCT

[0141] GGAGCTCCAGGTTTTCCAGGAGCTAGAGGTCCTTCTGGACCTCAAGGACCAT

[0142] CTGGTCCACCTGGACCAAAAGGTAACTCTGGAGAACCAGGTGCTCCTGGTTC

[0143] CAAAGGTGATACCGGTGCTAAAGGAGAGCCAGGTCCTACTGGTATTCAAGGT

[0144] CCTCCTGGTCCTGCTGGTGAAGAGGGTAAAAGAGGAGCCAGAGGTGAACCT

[0145] GGTCCAGCCGGTTTGCCAGGTCCACCAGGTGAAAGAGGAGGTCCAGGATCT

[0146] AGAGGTTTTCCAGGTGCTGACGGTGTTGCTGGTCCAAAGGGTCCTGCTGGTG

[0147] AGAGAGGAGCCCCAGGTCCAGCTGGACCTAAAGGATCTCCAGGAGAAGCTG

[0148] GTCGTCCTGGTGAAGCTGGTCTTCCAGGTGCTAAGGGTTTGACAGGCTCCCC

[0149] AGGATCCCCTGGTCCAGATGGTAAGACTGGACCACCTGGACCAGCTGGTCA

[0150] AGATGGAAGACCAGGTCCTCCAGGACCACCTGGAGCCAGAGGCCAAGCTGG

[0151] TGTTATGGGTTTCCCTGGTCCAAAAGGTGCTGCTGGAGAACCGGGTAAAGCC

[0152] GGTGAAAGAGGTGTTCCAGGTCCTCCAGGTGCTGTTGGTCCTGCTGGTAAAG

[0153] ATGGAGAAGCCGGTGCTCAAGGACCACCAGGTCCAGCTGGTCCAGCTGGAG

[0154] AGAGAGGTGAACAAGGTCCTGCTGGATCCCCAGGTTTCCAAGGTTTGCCTGG

[0155] TCCTGCTGGACCACCAGGTGAAGCTGGTAAGCCAGGTGAACAAGGAGTTCC

[0156] AGGTGATTTGGGTGCTCCTGGTCCATCCGGTGCTAGAGGTGAAAGAGGATTT

[0157] CCAGGAGAGAGAGGTGTTCAAGGACCACCTGGTCCTGCTGGACCAAGAGGA

[0158] GCAAACGGTGCTCCAGGAAACGATGGTGCTAAAGGTGATGCTGGTGCTCCA GGAGCTCCAGGTTCACAAGGTGCTCCAGGTTTGCAAGGAATGCCTGGAGAA AGAGGTGCTGCTGGATTACCAGGTCCAAAAGGAGACAGAGGTGATGCCGGT CCAAAGGGTGCTGATGGTGCTCCAGGTAAGGACGGTGTGCGAGGTTTGACT GGACCAATTGGACCTCCAGGTCCAGCTGGAGCCCCAGGAGATAAGGGTGAA

[0159] GCTGGTCCATCGGGTCCTGCTGGTCCTACTGGTGCTAGAGGAGCTCCAGGA GATAGAGGTGAACCTGGTCCACCTGGACCCGCTGGATTTGCTGGTCCACCA GGTGCTGATGGTCAACCAGGGGCTAAAGGTGAACCAGGTGACGCTGGAGCT AAAGGAGATGCCGGTCCACCAGGTCCAGCCGGTCCAGCTGGTCCTCCTGGT

[0160] CCAATTGGTAATGTTGGCGCTCCTGGTCCTAAGGGTGCCAGAGGTTCAGCTG GACCACCAGGTGCTACTGGTTTCCCAGGTGCTGCCGGTAGAGTTGGTCCAC CAGGTCCATCTGGTAACGCTGGTCCTCCAGGTCCACCAGGTCCTGCTGGAAA GGAGGGATCTAAGGGACCAAGAGGTGAAACTGGTCCAGCTGGAAGACCTGG

[0161] TGAAGTTGGTCCTCCAGGACCACCAGGTCCTGCTGGAGAAAAAGGTGCTCCA GGTGCTGATGGACCAGCTGGTGCTCCTGGTACCCCAGGTCCACAAGGTATT GCTGGTCAAAGAGGTGTGGTTGGTCTTCCTGGACAAAGAGGTGAGAGAGGTT TTCCTGGTTTGCCAGGTCCTTCTGGAGAACCTGGAAAACAGGGTCCCTCTGG

[0162] TGCTTCTGGTGAGAGAGGTCCTCCAGGACCAATGGGACCTCCAGGTTTGGCT GGTCCACCAGGTGAGTCTGGTAGAGAGGGTGCTCCAGGTGCTGAGGGTTCT CCAGGTAGAGATGGTAGTCCAGGTGCTAAAGGAGACAGAGGAGAGACTGGT CCAGCTGGTCCTCCAGGTGCTCCTGGTGCTCCCGGTGCTCCTGGTCCTGTC

[0163] GGTCCTGCTGGAAAGTCTGGTGATAGAGGTGAGACTGGTCCAGCTGGACCA GCTGGTCCAATCGGACCAGTTGGAGCTAGAGGTCCTGCTGGTCCTCAAGGT CCAAGAGGTGACAAGGGTGAAACAGGTGAGCAAGGTGACAGAGGTATTAAA GGACATAGAGGTTTCTCTGGTCTACAGGGACCTCCTGGTCCTCCAGGTTCCC

[0164] CAGGTGAGCAAGGTCCTTCTGGTGCTTCAGGTCCTGCTGGTCCAAGAGGTCC ACCAGGTTCTGCTGGTTCTCCAGGAAAGGATGGTTTGAATGGTTTGCCTGGA CCTATTGGTCCTCCTGGACCTAGAGGTAGAACAGGTGATGCTGGTCCTGCTG GTCCACCTGGTCCTCCTGGTCCTCCTGGTCCCCCTGGACCTCCATCTGGAGG

[0165] TTACGATTTGTCTTTTTTGCCACAACCTCCACAGGAAAAAGCCCATGACGGTG GAAGATATTATAGAGCTTAA

[0166] A corresponding collagen sequence may be as follows of SEQ ID NO: 6 as depicted below. In a preferred embodiment, the collagen that is expressed by the non-animal cell has at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% homology (or identity) of a collagen of SEQ ID NO: 6:

[0167] MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLEGDFDVAVLPFS

[0168] NSTNNGLLFINTTIASIAAKEEGVSLEKREAEAHHHHHHENLYFQGQLSYGYDEK STGISVPGPMGPSGPRGLPGPPGAPGPQGFQGPPGEPGEPGASGPMGPRGPP GPPGKNGDDGEAGKPGRPGERGPPGPQGARGLPGTAGLPGMKGHRGFSGLD GAKGDAGPAGPKGEPGSPGENGAPGQMGPRGLPGERGRPGAPGPAGARGND GATGAAGPPGPTGPAGPPGFPGAVGAKGEGGPQGPRGSEGPQGVRGEPGPP GPAGAAGPAGNPGADGQPGAKGANGAPGIAGAPGFPGARGPSGPQGPSGPP GPKGNSGEPGAPGSKGDTGAKGEPGPTGIQGPPGPAGEEGKRGARGEPGPAG LPGPPGERGGPGSRGFPGADGVAGPKGPAGERGAPGPAGPKGSPGEAGRPG EAGLPGAKGLTGSPGSPGPDGKTGPPGPAGQDGRPGPPGPPGARGQAGVMG FPGPKGAAGEPGKAGERGVPGPPGAVGPAGKDGEAGAQGPPGPAGPAGERG EQGPAGSPGFQGLPGPAGPPGEAGKPGEQGVPGDLGAPGPSGARGERGFPG ERGVQGPPGPAGPRGANGAPGNDGAKGDAGAPGAPGSQGAPGLQGMPGER GAAGLPGPKGDRGDAGPKGADGAPGKDGVRGLTGPIGPPGPAGAPGDKGEAG

[0169] PSGPAGPTGARGAPGDRGEPGPPGPAGFAGPPGADGQPGAKGEPGDAGAKG DAGPPGPAGPAGPPGPIGNVGAPGPKGARGSAGPPGATGFPGAAGRVGPPGP SGNAGPPGPPGPAGKEGSKGPRGETGPAGRPGEVGPPGPPGPAGEKGAPGA DGPAGAPGTPGPQGIAGQRGWGLPGQRGERGFPGLPGPSGEPGKQGPSGAS

[0170] GERGPPGPMGPPGLAGPPGESGREGAPGAEGSPGRDGSPGAKGDRGETGPA GPPGAPGAPGAPGPVGPAGKSGDRGETGPAGPAGPIGPVGARGPAGPQGPRG DKGETGEQGDRGIKGHRGFSGLQGPPGPPGSPGEQGPSGASGPAGPRGPPG SAGSPGKDGLNGLPGPIGPPGPRGRTGDAGPAGPPGPPGPPGPPGPPSGGYD

[0171] LSFLPQPPQEKAHDGGRYYRA

[0172] In a preferred embodiment, a further polypeptide that is expressed by the nonanimal cell (preferably in addition to one or more collagens) has at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% homology (or identity) of prolyl 4-hydroxylase subunit alpha-1 (Q1 RMLI3) of SEQ ID NO: 7:

[0173] HPGFFTSIGQMTDLIHTEKDLVTSLKDYIKAEEDKLEQIKKWAEKLDRLTSTATKD PEGFVGHPVNAFKLMKRLNTEWSELENLVLKDMSDGFISNLTIQRQYFPNDEDQ VGAAKALLRLQDTYNLDTDTISKGDLPGVKHKSFLTVEDCFELGKVAYTEADYYH TELWMEQALRQLDEGEVSTVDKVSVLDYLSYAVYQQGDLDKALLLTKKLLELDP

[0174] EHQRANGNLKYFEYIMAKEKDANKSSSDDQSDQKTTLKKKGAAVDYLPERQKYE MLCRGEGIKMTPRRQKKLFCRYHDGNRNPKFILAPAKQEDEWDKPRIIRFHDIIS DAEIEWKDLAKPRLRRATISNPITGDLETVHYRISKSAWLSGYENPVVSRINMRIQ DLTGLDVSTAEELQVANYGVGGQYEPHFDFARKDEPDAFKELGTGNRIATWLFY

[0175] M S D VLAG G ATVF P E VGAS VWP KKGTAVF WYN L FAS G E G D YSTR H AAC P VLVG N KWVSNKWLHERGQEFRRPCTLSELE In a preferred embodiment, the optional further polypeptide that is expressed by the non-animal cell (preferably in addition to one or more collagens) may be based on a DNA sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% homology (or identity) of codon-optimized polypeptide of SEQ ID NO: 8:

[0176] CATCCTGGTTTCTTTACTTCTATTGGACAAATGACTGACTTGATCCATACCGAA AAGGATTTGGTTACTTCCTTGAAAGATTACATTAAGGCTGAAGAGGATAAATT GGAACAGATTAAAAAATGGGCTGAAAAGTTGGATAGATTGACTTCCACAGCTA CTAAAGATCCAGAAGGTTTCGTTGGTCATCCAGTTAACGCTTTTAAGTTGATG AAGAGATTGAACACTGAATGGTCTGAGTTGGAAAATTTGGTCCTAAAGGATAT GTCTGACGGTTTTATCTCTAACTTGACTATCCAAAGACAATACTTCCCAAATGA TGAAGACCAAGTTGGTGCTGCTAAAGCTCTTTTGAGACTGCAAGACACCTAC AACTTGGATACCGATACAATTTCTAAGGGTGATTTGCCAGGTGTTAAGCACAA GTCTTTTTTAACCGTTGAAGATTGTTTCGAGTTGGGAAAGGTTGCATATACTG AGGCTGACTACTACCATACTGAATTGTGGATGGAACAGGCTTTGAGACAATTA GATGAAGGTGAGGTTTCTACAGTTGATAAGGTTTCTGTTTTGGATTACTTGTC TTATGCAGTTTACCAGCAAGGTGACTTGGATAAAGCATTGTTGTTGACTAAGA AGTTGTTGGAATTGGACCCTGAGCATCAAAGAGCAAATGGAAACTTGAAATAT TTTGAGTACATTATGGCTAAAGAAAAAGATGCAAACAAGTCTTCTTCTGATGAT CAATCCGACCAAAAGACAACTTTGAAGAAGAAGGGTGCTGCTGTTGATTACC TTCCAGAAAGACAGAAGTACGAGATGTTATGTAGAGGTGAAGGAATTAAGAT GACTCCAAGAAGACAGAAAAAGTTGTTTTGTAGATACCATGACGGTAACAGAA ACCCAAAGTTTATTTTGGCTCCTGCTAAGCAGGAGGATGAGTGGGATAAGCC TAGAATTATCAGATTCCATGACATTATCTCTGACGCTGAGATCGAGGTTGTTA AGGATTTGGCTAAGCCAAGATTGAGAAGAGCTACTATTTCTAATCCTATCACT GGTGATTTGGAGACTGTTCATTATAGAATTTCTAAATCCGCTTGGTTGTCTGG TTATGAAAACCCTGTTGTGTCTAGAATTAACATGAGAATTCAAGACCTCACTG GTTTGGATGTTTCTACAGCCGAAGAATTACAAGTCGCTAACTACGGTGTTGGT GGACAATACGAGCCACATTTCGACTTTGCTCGTAAGGATGAACCAGACGCTT TTAAGGAATTAGGTACCGGTAACAGAATCGCTACATGGCTTTTCTACATGTCT GATGTTCTGGCTGGAGGAGCTACTGTTTTTCCAGAAGTTGGTGCTTCTGTCTG GCCAAAGAAAGGTACCGCTGTGTTTTGGTATAATTTGTTCGCATCTGGTGAAG GCGATTACTCTACCAGACATGCTGCTTGTCCAGTCTTGGTGGGTAATAAGTG GGTTTCCAACAAATGGTTGCATGAAAGAGGTCAGGAGTTCAGAAGGCCATGT ACCTTGTCTGAATTGGAATAA In a preferred embodiment, a further polypeptide that is expressed by the nonanimal cell (preferably in addition to one or more collagens) has at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% homology (or identity) of protein disulfide-isomerase or prolyl 4-hydroxylase subunit beta (P05307, PDIA1_BOVIN) of SEQ ID NO: 9:

[0177] APDEEDHVLVLHKGNFDEALAAHKYLLVEFYAPWCGHCKALAPEYAKAAGKLKA EGSEIRLAKVDATEESDLAQQYGVRGYPTIKFFKNGDTASPKEYTAGREADDIVN WLKKRTGPAASTLSDGAAAEALVESSEVAVIGFFKDMESDSAKQFFLAAEVIDDI PFGITSNSDVFSKYQLDKDGWLFKKFDEGRNNFEGEVTKEKLLDFIKHNQLPLVI EFTEQTAPKIFGGEIKTHILLFLPKSVSDYEGKLSNFKKAAESFKGKILFIFIDSDHT DNQRILEFFGLKKEECPAVRLITLEEEMTKYKPESDELTAEKITEFCHRFLEGKIKP HLMSQELPDDWDKQPVKVLVGKNFEEVAFDEKKNVFVEFYAPWCGHCKQLAPI WDKLGETYKDHENIVIAKMDSTANEVEAVKVHSFPTLKFFPASADRTVIDYNGER TLDGFKKFLESGGQDGAGDDDDLEDLEEAEEPDLEEDDDQKAVKDEL

[0178] Bovine Collagen alpha-1 (I) Expression vector 1 ;

[0179] SEQ ID NO: 10:

[0180] CAACTTTGTATAGAAAAGTTGGCCAACATGGTGGAGCACGACACTCTCGTCT

[0181] ACTCCAAGAATATCAAAGATACAGTCTCAGAAGACCAAAGGGCTATTGAGACT

[0182] TTTCAACAAAGGGTAATATCGGGAAACCTCCTCGGATTCCATTGCCCAGCTAT CTGTCACTTCATCAAAAGGACAGTAGAAAAGGAAGGTGGCACCTACAAATGC CATCATTGCGATAAAGGAAAGGCTATCGTTCAAGATGCCTCTGCCGACAGTG GTCCCAAAGATGGACCCCCACCCACGAGGAGCATCGTGGAAAAAGAAGACG TTCCAACCACGTCTTCAAAGCAAGTGGATTGATGTGAACATGGTGGAGCACG ACACTCTCGTCTACTCCAAGAATATCAAAGATACAGTCTCAGAAGACCAAAGG GCTATTGAGACTTTTCAACAAAGGGTAATATCGGGAAACCTCCTCGGATTCCA TTGCCCAGCTATCTGTCACTTCATCAAAAGGACAGTAGAAAAGGAAGGTGGC ACCTACAAATGCCATCATTGCGATAAAGGAAAGGCTATCGTTCAAGATGCCTC TGCCGACAGTGGTCCCAAAGATGGACCCCCACCCACGAGGAGCATCGTGGA AAAAGAAGACGTTCCAACCACGTCTTCAAAGCAAGTGGATTGATGTGATATCT CCACTGACGTAAGGGATGACGCACAATCCCACTATCCTTCGCAAGACCCTTC CTCTATATAAGGAAGTTCATTTCATTTGGAGAGGACACGCTGACAAGTTTGTA CAAAAAAGCAGGCTGCCACCATGCATCACCATCATCATCATGGATCTGAGAA TTTATATTTTCAAGGTAGCATGTTTTCTTTCGTTGATTTGAGGTTGCTCCTTCTT TTAGCAGCTACTGCATTGCTTACTCATGGTCAAGAAGAGGGTCAAGAGGAAG GCCAAGAGGAGGATATTCCACCAGTCACTTGCGTGCAAAATGGTCTTAGGTA TCATGATAGAGATGTCTGGAAACCTGTTCCATGTCAAATTTGTGTGTGCGATA ATGGAAACGTGCTATGCGATGATGTTATTTGCGATGAATTGAAAGATTGCCCA

[0183] AATGCTAAGGTGCCAACTGATGAATGTTGTCCAGTTTGCCCAGAGGGACAAG

[0184] AAAGTCCAACAGATCAAGAAACTACAGGTGTTGAAGGTCCAAAGGGGGATAC

[0185] TGGCCCTAGAGGTCCACGGGGTCCAGCTGGACCACCAGGAAGAGATGGAAT

[0186] TCCAGGTCAACCTGGACTTCCAGGTCCACCAGGACCACCAGGCCCACCAGG

[0187] TCCTCCTGGTTTGGGAGGTAATTTTGCACCACAATTGAGTTATGGATACGACG

[0188] AAAAATCAACTGGAATATCAGTGCCAGGACCAATGGGTCCATCAGGACCTAG

[0189] GGGTCTTCCAGGGCCACCTGGCGCTCCTGGTCCTCAGGGATTTCAAGGTCC

[0190] TCCAGGTGAACCTGGAGAGCCTGGTGCTTCTGGTCCAATGGGACCAAGAGG

[0191] CCCTCCTGGTCCACCTGGTAAAAATGGTGATGATGGTGAGGCTGGAAAGCCT

[0192] GGTCGTCCTGGAGAAAGAGGACCACCAGGACCACAGGGAGCTAGAGGTTTG

[0193] CCAGGTACAGCCGGTCTTCCAGGAATGAAAGGACACAGGGGTTTTTCTGGTC

[0194] TTGATGGTGCTAAGGGAGACGCAGGACCTGCTGGACCAAAGGGTGAACCTG

[0195] GATCTCCTGGTGAAAATGGTGCTCCTGGACAAATGGGACCTAGAGGATTGCC

[0196] TGGCGAAAGAGGAAGACCTGGTGCTCCTGGACCAGCTGGAGCTAGAGGAAA

[0197] TGATGGAGCTACTGGTGCTGCCGGACCACCAGGACCAACTGGACCAGCTGG

[0198] ACCACCTGGTTTTCCAGGAGCAGTTGGGGCTAAGGGAGAAGGTGGACCACA

[0199] AGGACCTAGAGGATCAGAAGGACCTCAAGGAGTTAGGGGAGAACCAGGTCC

[0200] TCCTGGTCCTGCTGGAGCTGCTGGACCCGCTGGTAACCCAGGAGCAGATGG

[0201] ACAACCTGGAGCAAAAGGGGCTAATGGTGCTCCTGGTATTGCAGGTGCTCCT

[0202] GGATTCCCTGGTGCTAGAGGACCTTCAGGACCTCAGGGACCATCCGGTCCT

[0203] CCAGGACCAAAGGGAAACTCTGGTGAACCAGGGGCTCCTGGATCTAAGGGA

[0204] GATACTGGAGCTAAGGGTGAGCCAGGACCTACAGGAATTCAAGGACCACCA

[0205] GGACCTGCAGGAGAGGAAGGTAAGAGGGGTGCTAGAGGTGAACCTGGACCA

[0206] GCTGGATTGCCAGGCCCACCTGGAGAGAGGGGTGGTCCAGGTTCTAGAGGT

[0207] >

[0208] TTTCCAGGAGCTGATGGAGTTGCTGGTCCAAAGGGTCCTGCTGGTGAGAGA

[0209] GGTGCACCTGGACCAGCAGGACCTAAAGGATCACCTGGAGAAGCTGGTCGT

[0210] CCTGGTGAGGCTGGTCTTCCAGGTGCTAAGGGCCTTACAGGATCTCCAGGAT

[0211] CACCAGGTCCAGATGGAAAGACTGGACCACCAGGTCCTGCAGGACAAGATG

[0212] GAAGGCCTGGTCCACCTGGACCACCTGGTGCTAGGGGACAAGCTGGTGTTA

[0213] TGGGATTCCCAGGTCCAAAGGGTGCAGCTGGTGAGCCTGGTAAAGCTGGTG

[0214] AAAGAGGAGTTCCAGGACCTCCTGGTGCTGTTGGACCTGCTGGTAAAGATGG

[0215] TGAAGCAGGTGCTCAGGGTCCACCAGGTCCAGCCGGTCCAGCTGGTGAGAG

[0216] GGGAGAGCAAGGTCCTGCTGGATCTCCTGGATTTCAAGGTTTGCCTGGACCT

[0217] GCTGGACCTCCAGGAGAGGCTGGTAAACCTGGTGAGCAGGGTGTGCCTGGT

[0218] GATCTTGGTGCTCCTGGACCTAGTGGCGCAAGAGGCGAAAGGGGATTTCCA

[0219] GGTGAAAGAGGAGTTCAGGGACCACCAGGTCCAGCTGGTCCAAGAGGCGCT

[0220] AATGGAGCTCCAGGAAATGATGGAGCAAAGGGTGATGCTGGCGCTCCAGGA GCTCCTGGTTCTCAAGGTGCACCTGGATTGCAAGGAATGCCAGGAGAGAGA

[0221] GGTGCAGCTGGTTTGCCGGGACCAAAGGGTGATAGAGGTGATGCTGGTCCT

[0222] AAGGGAGCTGATGGAGCTCCAGGAAAGGATGGTGTGAGAGGATTGACTGGT

[0223] CCAATTGGACCACCAGGTCCTGCTGGAGCACCAGGAGATAAAGGAGAAGCT

[0224] GGTCCTTCAGGACCAGCCGGTCCAACTGGAGCTAGAGGAGCTCCAGGAGAT

[0225] AGAGGTGAACCAGGACCACCAGGACCTGCTGGGTTTGCAGGTCCACCTGGT

[0226] GCTGATGGACAACCTGGTGCTAAAGGAGAACCAGGCGATGCTGGTGCTAAG

[0227] GGAGATGCTGGCCCACCTGGTCCAGCTGGTCCTGCTGGGCCACCAGGTCCT

[0228] ATCGGAAATGTTGGTGCTCCAGGGCCTAAAGGAGCTAGAGGATCAGCAGGT

[0229] CCCCCAGGTGCTACAGGTTTTCCAGGGGCTGCTGGAAGAGTGGGTCCCCCA

[0230] GGTCCTTCTGGGAATGCAGGACCACCAGGTCCTCCAGGACCAGCTGGAAAA

[0231] GAAGGAAGTAAGGGACCAAGGGGAGAAACTGGACCTGCAGGACGTCCAGGT

[0232] GAAGTTGGTCCACCTGGACCTCCAGGTCCAGCTGGAGAGAAAGGAGCTCCA

[0233] GGAGCTGATGGTCCAGCAGGTGCTCCTGGTACTCCAGGTCCACAAGGTATTG

[0234] CTGGTCAAAGGGGAGTGGTTGGTCTCCCAGGTCAGAGAGGTGAAAGAGGTT

[0235] TCCCAGGTTTGCCTGGACCATCTGGAGAACCAGGTAAGCAGGGCCCATCTG

[0236] GAGCTTCTGGTGAGAGAGGGCCACCTGGACCTATGGGACCTCCTGGGCTTG

[0237] CTGGACCTCCAGGGGAATCTGGTAGGGAGGGTGCTCCAGGTGCTGAAGGAT

[0238] CACCTGGTAGAGATGGATCACCTGGAGCTAAAGGTGATAGAGGGGAAACAG

[0239] GTCCAGCAGGTCCACCTGGCGCTCCTGGTGCTCCAGGAGCACCAGGGCCAG

[0240] TTGGACCTGCTGGAAAGTCTGGAGATAGGGGAGAGACTGGACCAGCTGGAC

[0241] CTGCTGGACCAATCGGTCCTGTTGGCGCTAGAGGTCCTGCTGGACCACAAG

[0242] GTCCTAGGGGAGATAAAGGTGAAACAGGTGAACAAGGAGATAGAGGAATTAA

[0243] AGGTCATAGAGGGTTTTCTGGTTTGCAAGGACCTCCAGGACCTCCTGGATCT

[0244] CCAGGAGAGCAAGGACCATCTGGTGCAAGCGGACCTGCTGGTCCTAGAGGA

[0245] CCTCCAGGTTCAGCTGGCTCACCTGGTAAGGATGGATTAAATGGTTTGCCTG

[0246] GACCAATAGGTCCACCAGGACCTAGAGGAAGGACAGGTGATGCAGGACCAG

[0247] CTGGTCCACCTGGTCCACCAGGACCACCAGGACCACCAGGTCCACCTTCTG

[0248] GTGGATATGATCTTAGTTTTCTTCCTCAACCTCCCCAGGAGAAAGCTCATGAT

[0249] GGTGGTAGATATTATAGGGCTGATGATGCAAATGTTGTTAGAGATAGAGATCT

[0250] TGAGGTTGATACTACTCTTAAATCTCTTAGCCAACAAATTGAAAACATTCGTTC

[0251] TCCTGAGGGAAGCAGAAAAAATCCTGCAAGGACTTGTAGAGATCTTAAGATG

[0252] TGCCATTCAGATTGGAAGTCTGGTGAGTACTGGATTGATCCAAATCAAGGTTG

[0253] TAATTTGGATGCTATCAAAGTTTTTTGTAATATGGAAACTGGAGAGACATGTGT

[0254] GTACCCTACACAACCTTCTGTTGCTCAAAAAAATTGGTATATTTCAAAAAATCC

[0255] TAAAGAAAAGAGACATGTTTGGTATGGAGAATCTATGACTGGAGGATTTCAAT

[0256] TTGAGTATGGTGGACAAGGATCAGATCCAGCTGATGTGGCTATTCAACTTACT

[0257] TTTCTTAGATTGATGTCCACTGAAGCTTCTCAAAATATTACATATCATTGTAAG AATTCTGTTGCTTATATGGATCAACAGACTGGAAATCTTAAGAAGGCATTGTT

[0258] GTTGCAAGGCTCAAACGAAATTGAGATTAGGGCTGAAGGTAATAGTCGATTTA

[0259] CTTATTCTGTTACATATGATGGATGTACATCTCACACAGGTGCTTGGGGAAAA

[0260] ACAGTTATTGAGTATAAAACTACAAAAACTAGTAGACTTCCAATTATTGATGTT

[0261] GCACCATTGGATGTTGGTGCTCCTGATCAAGAATTTGGGTTTGATGTTGGACC

[0262] TGCTTGTTTCCTTGCATTTGTTTATTAAACCCAGCTTTCTTGTACAAAGTGGTA

[0263] GCTCGAATTTCCCCGATCGTTCAAACATTTGGCAATAAAGTTTCTTAAGATTGA

[0264] ATCCTGTTGCCGGTCTTGCGATGATTATCATATAATTTCTGTTGAATTACGTTA

[0265] AGCATGTAATAATTAACATGTAATGCATGACGTTATTTATGAGATGGGTTTTTA

[0266] TGATTAGAGTCCCGCAATTATACATTTAATACGCGATAGAAAACAAAATATAG

[0267] CGCGCAAACTAGGATAAATTATCGCGCGCGGTGTCATCTATGTTACTAGATC

[0268] GGGAAAGCTTTTAAACTATCAGTGTTTGACAGGATATATTGGCGGGTAAACCT

[0269] AAGAGAAAAGAGCGTTTATTAGAATAATCGGATATTTAAAAGGGCGTGAAAAG

[0270] GTTTATCCGTTCGTCCATTTGTATGTGCATGCCAACCACAGGGTTCCCCTCGG

[0271] GATCAAAGTACTTTGATCCAACCCCTCCGCTGCTATAGTGCAGTCGGCTTCTG

[0272] ACGTTCAGTGCAGCCGTCTTCTGAAAACGACATGTCGCACAAGTCCTAAGTTA

[0273] CGCGACAGGCTGCCGCCCTGCCCTTTTCCTGGCGTTTTCTTGTCGCGTGTTT

[0274] TAGTCGCATAAAGTAGAATACTTGCGACTAGAACCGGAGACATTACGCCATG

[0275] AACAAGAGCGCCGCCGCTGGCCTGCTGGGCTATGCCCGCGTCAGCACCGAC

[0276] GACCAGGACTTGACCAACCAACGGGCCGAACTGCACGCGGCCGGCTGCACC

[0277] AAGCTGTTTTCCGAGAAGATCACCGGCACCAGGCGCGACCGCCCGGAGCTG

[0278] GCCAGGATGCTTGACCACCTACGCCCTGGCGACGTTGTGACAGTGACCAGG

[0279] CTAGACCGCCTGGCCCGCAGCACCCGCGACCTACTGGACATTGCCGAGCGC

[0280] ATCCAGGAGGCCGGCGCGGGCCTGCGTAGCCTGGCAGAGCCGTGGGCCGA

[0281] CACCACCACGCCGGCCGGCCGCATGGTGTTGACCGTGTTCGCCGGCATTGC

[0282] CGAGTTCGAGCGTTCCCTAATCATCGACCGCACCCGGAGCGGGCGCGAGGC

[0283] CGCCAAGGCCCGAGGCGTGAAGTTTGGCCCCCGCCCTACCCTCACCCCGGC

[0284] ACAGATCGCGCACGCCCGCGAGCTGATCGACCAGGAAGGCCGCACCGTGAA

[0285] AGAGGCGGCTGCACTGCTTGGCGTGCATCGCTCGACCCTGTACCGCGCACT

[0286] TGAGCGCAGCGAGGAAGTGACGCCCACCGAGGCCAGGCGGCGCGGTGCCT

[0287] TCCGTGAGGACGCATTGACCGAGGCCGACGCCCTGGCGGCCGCCGAGAAT

[0288] GAACGCCAAGAGGAACAAGCATGAAACCGCACCAGGACGGCCAGGACGAAC

[0289] CGTTTTTCATTACCGAAGAGATCGAGGCGGAGATGATCGCGGCCGGGTACGT

[0290] GTTCGAGCCGCCCGCGCACGTCTCAACCGTGCGGCTGCATGAAATCCTGGC

[0291] CGGTTTGTCTGATGCCAAGCTGGCGGCCTGGCCGGCCAGCTTGGCCGCTGA

[0292] AGAAACCGAGCGCCGCCGTCTAAAAAGGTGATGTGTATTTGAGTAAAACAGC

[0293] TTGCGTCATGCGGTCGCTGCGTATATGATGCGATGAGTAAATAAACAAATAC

[0294] GCAAGGGGAACGCATGAAGGTTATCGCTGTACTTAACCAGAAAGGCGGGTC AGGCAAGACGACCATCGCAACCCATCTAGCCCGCGCCCTGCAACTCGCCGG

[0295] GGCCGATGTTCTGTTAGTCGATTCCGATCCCCAGGGCAGTGCCCGCGATTG

[0296] GGCGGCCGTGCGGGAAGATCAACCGCTAACCGTTGTCGGCATCGACCGCCC

[0297] GACGATTGACCGCGACGTGAAGGCCATCGGCCGGCGCGACTTCGTAGTGAT

[0298] CGACGGAGCGCCCCAGGCGGCGGACTTGGCTGTGTCCGCGATCAAGGCAG

[0299] CCGACTTCGTGCTGATTCCGGTGCAGCCAAGCCCTTACGACATATGGGCCAC

[0300] CGCCGACCTGGTGGAGCTGGTTAAGCAGCGCATTGAGGTCACGGATGGAAG

[0301] GCTACAAGCGGCCTTTGTCGTGTCGCGGGCGATCAAAGGCACGCGCATCGG

[0302] CGGTGAGGTTGCCGAGGCGCTGGCCGGGTACGAGCTGCCCATTCTTGAGTC

[0303] CCGTATCACGCAGCGCGTGAGCTACCCAGGCACTGCCGCCGCCGGCACAAC

[0304] CGTTCTTGAATCAGAACCCGAGGGCGACGCTGCCCGCGAGGTCCAGGCGCT

[0305] GGCCGCTGAAATTAAATCAAAACTCATTTGAGTTAATGAGGTAAAGAGAAAAT

[0306] GAGCAAAAGCACAAACACGCTAAGTGCCGGCCGTCCGAGCGCACGCAGCAG

[0307] CAAGGCTGCAACGTTGGCCAGCCTGGCAGACACGCCAGCCATGAAGCGGGT

[0308] CAACTTTCAGTTGCCGGCGGAGGATCACACCAAGCTGAAGATGTACGCGGTA

[0309] CGCCAAGGCAAGACCATTACCGAGCTGCTATCTGAATACATCGCGCAGCTAC

[0310] CAGAGTAAATGAGCAAATGAATAAATGAGTAGATGAATTTTAGCGGCTAAAGG

[0311] AGGCGGCATGGAAAATCAAGAACAACCAGGCACCGACGCCGTGGAATGCCC

[0312] CATGTGTGGAGGAACGGGCGGTTGGCCAGGCGTAAGCGGCTGGGTTGCCT

[0313] GCCGGCCCTGCAATGGCACTGGAACCCCCAAGCCCGAGGAATCGGCGTGA

[0314] GCGGTCGCAAACCATCCGGCCCGGTACAAATCGGCGCGGCGCTGGGTGATG

[0315] ACCTGGTGGAGAAGTTGAAGGCCGCGCAGGCCGCCCAGCGGCAACGCATC

[0316] GAGGCAGAAGCACGCCCCGGTGAATCGTGGCAAGCGGCCGCTGATCGAATC

[0317] CGCAAAGAATCCCGGCAACCGCCGGCAGCCGGTGCGCCGTCGATTAGGAAG

[0318] CCGCCCAAGGGCGACGAGCAACCAGATTTTTTCGTTCCGATGCTCTATGACG

[0319] TGGGCACCCGCGATAGTCGCAGCATCATGGACGTGGCCGTTTTCCGTCTGTC

[0320] GAAGCGTGACCGACGAGCTGGCGAGGTGATCCGCTACGAGCTTCCAGACGG

[0321] GCACGTAGAGGTTTCCGCAGGGCCGGCCGGCATGGCCAGTGTGTGGGATTA

[0322] CGACCTGGTACTGATGGCGGTTTCCCATCTAACCGAATCCATGAACCGATAC

[0323] CGGGAAGGGAAGGGAGACAAGCCCGGCCGCGTGTTCCGTCCACACGTTGC

[0324] GGACGTACTCAAGTTCTGCCGGCGAGCCGATGGCGGAAAGCAGAAAGACGA

[0325] CCTGGTAGAAACCTGCATTCGGTTAAACACCACGCACGTTGCCATGCAGCGT

[0326] ACGAAGAAGGCCAAGAACGGCCGCCTGGTGACGGTATCCGAGGGTGAAGCC

[0327] TTGATTAGCCGCTACAAGATCGTAAAGAGCGAAACCGGGCGGCCGGAGTAC

[0328] ATCGAGATCGAGCTAGCTGATTGGATGTACCGCGAGATCACAGAAGGCAAGA

[0329] ACCCGGACGTGCTGACGGTTCACCCCGATTACTTTTTGATCGATCCCGGCAT

[0330] CGGCCGTTTTCTCTACCGCCTGGCACGCCGCGCCGCAGGCAAGGCAGAAGC

[0331] CAGATGGTTGTTCAAGACGATCTACGAACGCAGTGGCAGCGCCGGAGAGTT CAAGAAGTTCTGTTTCACCGTGCGCAAGCTGATCGGGTCAAATGACCTGCCG GAGTACGATTTGAAGGAGGAGGCGGGGCAGGCTGGCCCGATCCTAGTCATG CGCTACCGCAACCTGATCGAGGGCGAAGCATCCGCCGGTTCCTAATGTACG GAGCAGATGCTAGGGCAAATTGCCCTAGCAGGGGAAAAAGGTCGAAAAGGT CTCTTTCCTGTGGATAGCACGTACATTGGGAACCCAAAGCCGTACATTGGGA ACCGGAACCCGTACATTGGGAACCCAAAGCCGTACATTGGGAACCGGTCAC ACATGTAAGTGACTGATATAAAAGAGAAAAAAGGCGATTTTTCCGCCTAAAAC TCTTTAAAACTTATTAAAACTCTTAAAACCCGCCTGGCCTGTGCATAACTGTCT GGCCAGCGCACAGCCGAAGAGCTGCAAAAAGCGCCTACCCTTCGGTCGCTG CGCTCCCTACGCCCCGCCGCTTCGCGTCGGCCTATCGCGGCCGCTGGCCG

[0332] CTCAAAAATGGCTGGCCTACGGCCAGGCAATCTACCAGGGCGCGGACAAGC CGCGCCGTCGCCACTCGACCGCCGGCGCCCACATCAAGGCACCCTGCCTCG CGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCGGAGAC GGTCACAGCTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGCCCGTCAGGG CGCGTCAGCGGGTGTTGGCGGGTGTCGGGGCGCAGCCATGACCCAGTCAC GTAGCGATAGCGGAGTGTATACTGGCTTAACTATGCGGCATCAGAGCAGATT GTACTGAGAGTGCACCATATGCGGTGTGAAATACCGCACAGATGCGTAAGGA GAAAATACCGCATCAGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCG CTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAAT ACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAA

[0333] GGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTC CATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGA GGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAA GCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTC CGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGG TATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAAC CCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTC CAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAG GATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGG CCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAA

[0334] GCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACC ACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAA AAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAG TGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGCATTCTAGGTACTAAAA CAATTCATCCAGTAAAATATAATATTTTATTTTCTCCCAATCAGGCTTGATCCC CAGTAAGTCAAAAAATAGCTCGACATACTGTTCTTCCCCGATATCCTCCCTGA TCGACCGGACGCAGAAGGCAATGTCATACCACTTGTCCGCCCTGCCGCTTCT CCCAAGATCAATAAAGCCACTTACTTTGCCATCTTTCACAAAGATGTTGCTGT CTCCCAGGTCGCCGTGGGAAAAGACAAGTTCCTCTTCGGGCTTTTCCGTCTT

[0335] TAAAAAATCATACAGCTCGCGCGGATCTTTAAATGGAGTGTCTTCTTCCCAGT

[0336] TTTCGCAATCCACATCGGCCAGATCGTTATTCAGTAAGTAATCCAATTCGGCT

[0337] AAGCGGCTGTCTAAGCTATTCGTATAGGGACAATCCGATATGTCGATGGAGT

[0338] GAAAGAGCCTGATGCACTCCGCATACAGCTCGATAATCTTTTCAGGGCTTTGT

[0339] TCATCTTCATACTCTTCCGAGCAAAGGACGCCATCGGCCTCACTCATGAGCA

[0340] GATTGCTCCAGCCATCATGCCGTTCAAAGTGCAGGACCTTTGGAACAGGCAG

[0341] CTTTCCTTCCAGCCATAGCATCATGTCCTTTTCCCGTTCCACATCATAGGTGG

[0342] TCCCTTTATACCGGCTGTCCGTCATTTTTAAATATAGGTTTTCATTTTCTCCCA

[0343] CCAGCTTATATACCTTAGCAGGAGACATTCCTTCCGTATCTTTTACGCAGCGG

[0344] TATTTTTCGATCAGTTTTTTCAATTCCGGTGATATTCTCATTTTAGCCATTTATT

[0345] ATTTCCTTCCTCTTTTCTACAGTATTTAAAGATACCCCAAGAAGCTAATTATAA

[0346] CAAGACGAACTCCAATTCACTGTTCCTTGCATTCTAAAACCTTAAATACCAGA

[0347] AAACAGCTTTTTCAAAGTTGTTTTCAAAGTTGGCGTATAACATAGTATCGACG

[0348] GAGCCGATTTTGAAACCGCGGTGATCACAGGCAGCAACGCTCTGTCATCGTT

[0349] ACAATCAACATGCTACCCTCCGCGAGATCATCCGTGTTTCAAACCCGGCAGC

[0350] TTAGTTGCCGTTCTTCCGAATAGCATCGGTAACATGAGCAAAGTCTGCCGCCT

[0351] TACAACGGCTCTCCCGCTGACGCCGTCCCGGACTGATGGGCTGCCTGTATC

[0352] GAGTGGTGATTTTGTGCCGAGCTGCCGGTCGGGGAGCTGTTGGCTGGCTGG

[0353] TGGCAGGATATATTGTGGTGTAAACAAATTGACGCTTAGACAACTTAATAACA

[0354] CATTGCGGACGTTTTTAATGTACTGAATTAACGCCGAATTGAATTCGAGCTCG

[0355] GTACCGGGGGATCTGGATTTTAGTACTGGATTTTGGTTTTAGGAATTAGAAAT

[0356] TTTATTGATAGAAGTATTTTACAAATACAAATACATACTAAGGGTTTCTTATATG

[0357] CTCAACACATGAGCGAAACCCTATAGGAACCCTAATTCCCTTATCTGGGAACT

[0358] ACTCACACATTATTATGGAGAAACTCGAGCTTGTCGATCGACTCAAATCTCGG

[0359] TGACGGGCAGGACCGGACGGGGCGGTACCGGCAGGCTGAAGTCCAGCTGC

[0360] CAGAAACCCACGTCATGCCAGTTCCCGTGCTTGAAGCCGGCCGCCCGCAGC

[0361] ATGCCGCGGGGGGCATATCCGAGCGCCTCGTGCATGCGCACGCTCGGGTC

[0362] GTTGGGCAGCCCGATGACAGCGACCACGCTCTTGAAGCCCTGTGCCTCCAG

[0363] GGACTTCAGCAGGTGGGTGTAGAGCGTGGAGCCCAGTCCCGTCCGCTGGTG

[0364] GCGGGGGGAGACGTACACGGTCGACTCGGCCGTCCAGTCGTAGGCGTTGC

[0365] GTGCCTTCCAGGGGCCCGCGTAGGCGATGCCGGCGACCTCGCCGTCCACCT

[0366] CGGCGACGAGCCAGGGATAGCGCTCCCGCAGACGGACGAGGTCGTCCGTC

[0367] CACTCCTGCGGTTCCTGCGGCTCGGTACGGAAGTTGACCGTGCTTGTCTCGA

[0368] TGTAGTGGTTGACGATGGTGCAGACCGCCGGCATGTCCGCCTCGGTGGCAC

[0369] GGCGGATGTCGGCCGGGCGTCGTTCTGGGCTCATGGTAGACGATCTGCGAA

[0370] AGCTCGAGAGAGATAGATTTGTAGAGAGAGACTGGTGATTTCAGCGTGTCCT

[0371] CTCCAAATGAAATGAACTTCCTTATATAGAGGAAGGGTCTTGCGAAGGATAGT GGGATTGTGCGTCATCCCTTACGTCAGTGGAGATATCACATCAATCCACTTGC TTTGAAGACGTGGTTGGAACGTCTTCTTTTTCCACGATGCTCCTCGTGGGTG GGGGTCCATCTTTGGGACCACTGTCGGCAGAGGCATCTTGAACGATAGCCTT TCCTTTATCGCAATGATGGCATTTGTAGGTGCCACCTTCCTTTTCTACTGTCCT TTTGATGAAGTGACAGATAGCTGGGCAATGGAATCCGAGGAGGTTTCCCGAT ATTACCCTTTGTTGAAAAGTCTCAATAGCCCTTTGGTCTTCTGAGACTGTATCT TTGATATTCTTGGAGTAGACGAGAGTGTCGTGCTCCACCATGTTCACATCAAT CCACTTGCTTTGAAGACGTGGTTGGAACGTCTTCTTTTTCCACGATGCTCCTC GTGGGTGGGGGTCCATCTTTGGGACCACTGTCGGCAGAGGCATCTTGAACG ATAGCCTTTCCTTTATCGCAATGATGGCATTTGTAGGTGCCACCTTCCTTTTCT ACTGTCCTTTTGATGAAGTGACAGATAGCTGGGCAATGGAATCCGAGGAGGT TTCCCGATATTACCCTTTGTTGAAAAGTCTCAATAGCCCTTTGGTCTTCTGAGA CTGTATCTTTGATATTCTTGGAGTAGACGAGAGTGTCGTGCTCCACCATGTTG

[0372] GCGTACCCGGGGATCCTCTAGAGTCGACCTGCAGGCATGCAAGCT

[0373] Bovine Collagen alpha-1 (I) Expression vector 2;

[0374] SEQ ID NO: 11 :

[0375] CAACTTTGTATAGAAAAGTTGGCCAACATGGTGGAGCACGACACTCTCGTCT ACTCCAAGAATATCAAAGATACAGTCTCAGAAGACCAAAGGGCTATTGAGACT TTTCAACAAAGGGTAATATCGGGAAACCTCCTCGGATTCCATTGCCCAGCTAT CTGTCACTTCATCAAAAGGACAGTAGAAAAGGAAGGTGGCACCTACAAATGC CATCATTGCGATAAAGGAAAGGCTATCGTTCAAGATGCCTCTGCCGACAGTG GTCCCAAAGATGGACCCCCACCCACGAGGAGCATCGTGGAAAAAGAAGACG TTCCAACCACGTCTTCAAAGCAAGTGGATTGATGTGAACATGGTGGAGCACG ACACTCTCGTCTACTCCAAGAATATCAAAGATACAGTCTCAGAAGACCAAAGG GCTATTGAGACTTTTCAACAAAGGGTAATATCGGGAAACCTCCTCGGATTCCA TTGCCCAGCTATCTGTCACTTCATCAAAAGGACAGTAGAAAAGGAAGGTGGC ACCTACAAATGCCATCATTGCGATAAAGGAAAGGCTATCGTTCAAGATGCCTC TGCCGACAGTGGTCCCAAAGATGGACCCCCACCCACGAGGAGCATCGTGGA

[0376] AAAAGAAGACGTTCCAACCACGTCTTCAAAGCAAGTGGATTGATGTGATATCT CCACTGACGTAAGGGATGACGCACAATCCCACTATCCTTCGCAAGACCCTTC CTCTATATAAGGAAGTTCATTTCATTTGGAGAGGACACGCTGACAAGTTTGTA CAAAAAAGCAGGCTGCCACCATGTTTTCTTTTGTTGATCTTAGATTGCTCCTTC TTCTTGCTGCTACAGCACTTCTTACTCACGGGCAAGAAGAAGGTCAGGAAGA AGGTCAAGAAGAAGATATTCCACCAGTTACTTGTGTTCAAAACGGTCTTAGAT ACCATGATAGGGATGTTTGGAAGCCTGTGCCATGTCAAATTTGTGTTTGTGAT AATGGAAATGTTTTATGTGATGATGTAATTTGTGATGAACTTAAGGATTGCCCT AATGCTAAGGTACCAACTGATGAATGTTGTCCTGTGTGTCCAGAGGGTCAAG AATCTCCTACAGATCAAGAAACTACTGGAGTTGAAGGACCAAAAGGTGATACT

[0377] GGACCCAGGGGCCCTAGGGGTCCAGCAGGTCCTCCAGGAAGGGATGGAATT

[0378] CCTGGCCAGCCTGGTCTACCAGGACCACCGGGACCACCTGGCCCTCCTGGT

[0379] CCACCTGGACTTGGAGGTAATTTTGCACCTCAACTTTCTTACGGATACGATGA

[0380] GAAGTCTACAGGTATTTCTGTTCCTGGACCTATGGGTCCTTCAGGTCCAAGA

[0381] GGTCTTCCAGGCCCTCCAGGAGCTCCAGGCCCTCAAGGATTTCAAGGTCCTC

[0382] CTGGTGAACCAGGAGAACCAGGAGCATCAGGACCAATGGGACCCAGAGGAC

[0383] CTCCAGGTCCTCCTGGAAAAAATGGTGATGATGGCGAGGCTGGTAAGCCAG

[0384] GTAGACCTGGTGAAAGAGGGCCTCCAGGTCCACAAGGTGCTAGAGGACTAC

[0385] CAGGAACAGCTGGTCTTCCAGGAATGAAGGGTCATAGGGGATTTTCTGGTTT

[0386] GGATGGAGCAAAAGGTGATGCTGGACCAGCTGGTCCAAAAGGAGAACCAGG

[0387] ATCTCCTGGAGAGAATGGAGCTCCAGGACAAATGGGACCTAGAGGACTTCCA

[0388] GGTGAAAGAGGAAGACCAGGAGCACCAGGACCTGCTGGTGCTAGAGGAAAT

[0389] GATGGGGCTACTGGTGCTGCTGGACCACCTGGACCAACAGGACCTGCAGGA

[0390] CCACCTGGTTTTCCAGGTGCTGTTGGAGCTAAGGGAGAGGGAGGACCACAA

[0391] GGACCTCGTGGCTCAGAAGGTCCACAAGGTGTCAGAGGTGAGCCAGGTCCA

[0392] CCTGGACCTGCTGGAGCAGCTGGTCCTGCTGGAAATCCCGGTGCTGATGGA

[0393] CAGCCTGGAGCCAAAGGAGCAAACGGAGCTCCTGGAATTGCTGGTGCACCT

[0394] GGTTTCCCTGGTGCCAGAGGACCTTCAGGTCCTCAGGGGCCTTCTGGTCCA

[0395] CCAGGTCCTAAAGGAAACTCTGGAGAGCCTGGAGCTCCTGGTTCTAAAGGAG

[0396] ATACAGGCGCTAAGGGCGAACCTGGACCTACAGGAATCCAAGGACCTCCTG

[0397] GTCCAGCAGGTGAGGAAGGGAAGAGAGGAGCTAGAGGAGAGCCTGGACCT

[0398] GCTGGTCTTCCTGGCCCACCAGGAGAAAGAGGAGGTCCTGGATCTAGAGGA

[0399] TTCCCAGGAGCTGATGGAGTTGCCGGACCTAAAGGACCTGCTGGTGAAAGA

[0400] GGAGCTCCAGGTCCAGCAGGTCCTAAAGGATCACCAGGAGAAGCTGGTAGG

[0401] CCAGGTGAAGCTGGTTTGCCAGGAGCTAAAGGTCTTACAGGATCTCCTGGCT

[0402] CTCCAGGTCCTGATGGGAAGACTGGACCACCAGGTCCTGCTGGACAAGATG

[0403] GAAGGCCAGGTCCTCCAGGACCACCTGGAGCAAGAGGTCAAGCTGGTGTGA

[0404] TGGGTTTTCCAGGACCAAAGGGAGCTGCAGGAGAACCAGGTAAGGCTGGAG

[0405] AAAGGGGAGTACCTGGTCCTCCTGGTGCTGTTGGTCCAGCTGGTAAAGATGG

[0406] AGAAGCAGGAGCACAAGGTCCTCCAGGTCCTGCAGGACCTGCTGGAGAAAG

[0407] AGGTGAACAAGGACCTGCAGGATCACCAGGATTTCAAGGATTGCCTGGCCCT

[0408] GCAGGACCTCCAGGTGAAGCTGGTAAGCCAGGAGAGCAAGGTGTCCCAGGT

[0409] GATCTCGGGGCTCCTGGTCCATCAGGAGCAAGAGGAGAGAGAGGATTTCCA

[0410] GGAGAAAGAGGAGTTCAGGGACCACCAGGACCAGCAGGACCAAGAGGTGCT

[0411] AACGGCGCACCAGGAAATGACGGTGCAAAGGGGGATGCTGGTGCTCCAGGA

[0412] GCTCCTGGATCACAAGGAGCTCCAGGTTTGCAAGGCATGCCAGGAGAAAGA

[0413] GGAGCAGCAGGATTGCCTGGACCTAAGGGTGATAGAGGCGATGCAGGACCA AAAGGAGCTGATGGTGCTCCAGGTAAGGATGGAGTTCGTGGATTGACAGGT

[0414] CCAATTGGTCCACCAGGACCAGCTGGAGCTCCTGGAGATAAAGGCGAAGCT

[0415] GGACCATCAGGACCAGCTGGACCAACTGGTGCTAGAGGAGCTCCTGGAGAC

[0416] AGGGGAGAGCCTGGACCTCCTGGTCCTGCTGGTTTCGCTGGACCTCCTGGA

[0417] GCAGATGGACAGCCAGGTGCAAAGGGAGAGCCAGGTGATGCCGGAGCTAAA

[0418] GGGGATGCTGGTCCACCTGGTCCAGCAGGTCCAGCTGGTCCTCCTGGCCCT

[0419] ATAGGTAATGTTGGTGCTCCTGGTCCAAAAGGAGCTAGAGGGTCTGCTGGTC

[0420] CTCCAGGAGCTACAGGTTTTCCTGGAGCAGCTGGAAGAGTTGGTCCTCCAGG

[0421] TCCTTCAGGAAATGCTGGACCTCCTGGTCCACCAGGACCTGCTGGAAAGGAA

[0422] GGATCAAAAGGTCCAAGAGGTGAAACTGGACCTGCTGGTAGGCCAGGAGAA

[0423] GTGGGTCCACCAGGACCACCAGGACCTGCAGGAGAAAAGGGTGCTCCAGGA

[0424] GCTGATGGTCCTGCTGGTGCTCCAGGAACACCTGGACCACAAGGAATTGCTG

[0425] GTCAAAGAGGTGTTGTGGGTCTTCCAGGTCAAAGAGGTGAAAGAGGTTTCCC

[0426] TGGACTTCCTGGTCCATCAGGTGAACCTGGAAAGCAAGGTCCAAGTGGAGCT

[0427] TCTGGAGAAAGAGGTCCTCCAGGACCTATGGGTCCTCCTGGTTTGGCAGGTC

[0428] CTCCTGGAGAATCTGGTAGAGAAGGTGCACCTGGAGCAGAAGGTTCTCCAG

[0429] GTAGGGATGGATCTCCTGGAGCTAAAGGAGATAGGGGTGAAACAGGACCTG

[0430] CAGGACCACCTGGAGCACCTGGAGCTCCTGGTGCTCCAGGACCAGTTGGAC

[0431] CAGCAGGTAAGTCTGGAGATCGTGGTGAAACTGGACCAGCTGGACCAGCTG

[0432] GACCTATTGGTCCTGTTGGTGCTAGGGGACCAGCAGGCCCACAAGGACCTA

[0433] GAGGTGATAAAGGAGAAACTGGTGAACAGGGTGATAGAGGAATAAAAGGCC

[0434] ATAGAGGGTTCAGTGGACTCCAAGGACCTCCTGGTCCACCAGGATCTCCTGG

[0435] AGAGCAAGGACCTTCTGGTGCTTCTGGCCCAGCTGGTCCTAGAGGACCTCCA

[0436] GGTTCTGCAGGATCTCCTGGTAAAGATGGATTGAACGGTCTTCCAGGACCTA

[0437] TTGGTCCTCCTGGACCTAGGGGAAGGACTGGTGATGCTGGTCCAGCTGGAC

[0438] CACCAGGTCCACCTGGTCCTCCAGGACCACCAGGTCCACCAAGTGGTGGTT

[0439] ATGATCTTAGCTTTCTTCCTCAACCTCCTCAAGAGAAGGCCCACGATGGAGG

[0440] ACGTTATTATAGAGCTGATGATGCTAACGTTGTTAGAGATAGAGATTTGGAAG

[0441] TTGATACTACTTTGAAATCTCTTTCACAACAAATTGAAAATATTAGATCACCTG

[0442] AAGGATCAAGGAAGAACCCTGCTAGGACTTGTAGAGATCTTAAAATGTGCCA

[0443] TAGCGATTGGAAGTCTGGAGAGTATTGGATTGATCCAAATCAAGGATGCAAC

[0444] TTGGATGCTATTAAAGTTTTCTGTAACATGGAGACTGGAGAAACATGTGTTTA

[0445] CCCAACTCAACCAAGCGTTGCTCAAAAAAACTGGTACATCTCAAAAAATCCAA

[0446] AAGAAAAGAGGCATGTGTGGTATGGAGAATCTATGACTGGAGGATTCCAGTT

[0447] TGAATATGGAGGTCAAGGTTCTGATCCTGCTGATGTTGCTATTCAATTGACTT

[0448] TCTTGAGACTTATGTCTACAGAGGCATCACAAAACATTACTTATCATTGCAAG

[0449] AACTCAGTTGCTTATATGGATCAACAAACTGGAAATCTTAAAAAGGCTCTTCT

[0450] CTTGCAAGGATCTAATGAAATTGAGATAAGAGCTGAAGGAAACTCTAGATTTA CTTATTCTGTTACATACGATGGATGCACTTCACATACTGGTGCATGGGGTAAA

[0451] ACTGTAATTGAGTACAAGACTACTAAGACATCTCGTCTTCCTATTATTGATGTT

[0452] GCTCCACTTGATGTTGGTGCTCCAGACCAAGAATTTGGTTTTGATGTGGGTCC

[0453] TGCATGTTTTCTTGCTTTTGTTTATTAAACCCAGCTTTCTTGTACAAAGTGGTA

[0454] GCTCGAATTTCCCCGATCGTTCAAACATTTGGCAATAAAGTTTCTTAAGATTGA

[0455] ATCCTGTTGCCGGTCTTGCGATGATTATCATATAATTTCTGTTGAATTACGTTA

[0456] AGCATGTAATAATTAACATGTAATGCATGACGTTATTTATGAGATGGGTTTTTA

[0457] TGATTAGAGTCCCGCAATTATACATTTAATACGCGATAGAAAACAAAATATAG

[0458] CGCGCAAACTAGGATAAATTATCGCGCGCGGTGTCATCTATGTTACTAGATC

[0459] GGGAAAGCTTTTAAACTATCAGTGTTTGACAGGATATATTGGCGGGTAAACCT

[0460] AAGAGAAAAGAGCGTTTATTAGAATAATCGGATATTTAAAAGGGCGTGAAAAG

[0461] GTTTATCCGTTCGTCCATTTGTATGTGCATGCCAACCACAGGGTTCCCCTCGG

[0462] GATCAAAGTACTTTGATCCAACCCCTCCGCTGCTATAGTGCAGTCGGCTTCTG

[0463] ACGTTCAGTGCAGCCGTCTTCTGAAAACGACATGTCGCACAAGTCCTAAGTTA

[0464] CGCGACAGGCTGCCGCCCTGCCCTTTTCCTGGCGTTTTCTTGTCGCGTGTTT

[0465] TAGTCGCATAAAGTAGAATACTTGCGACTAGAACCGGAGACATTACGCCATG

[0466] AACAAGAGCGCCGCCGCTGGCCTGCTGGGCTATGCCCGCGTCAGCACCGAC

[0467] GACCAGGACTTGACCAACCAACGGGCCGAACTGCACGCGGCCGGCTGCACC

[0468] AAGCTGTTTTCCGAGAAGATCACCGGCACCAGGCGCGACCGCCCGGAGCTG

[0469] GCCAGGATGCTTGACCACCTACGCCCTGGCGACGTTGTGACAGTGACCAGG

[0470] CTAGACCGCCTGGCCCGCAGCACCCGCGACCTACTGGACATTGCCGAGCGC

[0471] ATCCAGGAGGCCGGCGCGGGCCTGCGTAGCCTGGCAGAGCCGTGGGCCGA

[0472] CACCACCACGCCGGCCGGCCGCATGGTGTTGACCGTGTTCGCCGGCATTGC

[0473] CGAGTTCGAGCGTTCCCTAATCATCGACCGCACCCGGAGCGGGCGCGAGGC

[0474] CGCCAAGGCCCGAGGCGTGAAGTTTGGCCCCCGCCCTACCCTCACCCCGGC

[0475] ACAGATCGCGCACGCCCGCGAGCTGATCGACCAGGAAGGCCGCACCGTGAA

[0476] AGAGGCGGCTGCACTGCTTGGCGTGCATCGCTCGACCCTGTACCGCGCACT

[0477] TGAGCGCAGCGAGGAAGTGACGCCCACCGAGGCCAGGCGGCGCGGTGCCT

[0478] TCCGTGAGGACGCATTGACCGAGGCCGACGCCCTGGCGGCCGCCGAGAAT

[0479] GAACGCCAAGAGGAACAAGCATGAAACCGCACCAGGACGGCCAGGACGAAC

[0480] CGTTTTTCATTACCGAAGAGATCGAGGCGGAGATGATCGCGGCCGGGTACGT

[0481] GTTCGAGCCGCCCGCGCACGTCTCAACCGTGCGGCTGCATGAAATCCTGGC

[0482] CGGTTTGTCTGATGCCAAGCTGGCGGCCTGGCCGGCCAGCTTGGCCGCTGA

[0483] AGAAACCGAGCGCCGCCGTCTAAAAAGGTGATGTGTATTTGAGTAAAACAGC

[0484] TTGCGTCATGCGGTCGCTGCGTATATGATGCGATGAGTAAATAAACAAATAC

[0485] GCAAGGGGAACGCATGAAGGTTATCGCTGTACTTAACCAGAAAGGCGGGTC

[0486] AGGCAAGACGACCATCGCAACCCATCTAGCCCGCGCCCTGCAACTCGCCGG

[0487] GGCCGATGTTCTGTTAGTCGATTCCGATCCCCAGGGCAGTGCCCGCGATTG GGCGGCCGTGCGGGAAGATCAACCGCTAACCGTTGTCGGCATCGACCGCCC

[0488] GACGATTGACCGCGACGTGAAGGCCATCGGCCGGCGCGACTTCGTAGTGAT

[0489] CGACGGAGCGCCCCAGGCGGCGGACTTGGCTGTGTCCGCGATCAAGGCAG

[0490] CCGACTTCGTGCTGATTCCGGTGCAGCCAAGCCCTTACGACATATGGGCCAC

[0491] CGCCGACCTGGTGGAGCTGGTTAAGCAGCGCATTGAGGTCACGGATGGAAG

[0492] GCTACAAGCGGCCTTTGTCGTGTCGCGGGCGATCAAAGGCACGCGCATCGG

[0493] CGGTGAGGTTGCCGAGGCGCTGGCCGGGTACGAGCTGCCCATTCTTGAGTC

[0494] CCGTATCACGCAGCGCGTGAGCTACCCAGGCACTGCCGCCGCCGGCACAAC

[0495] CGTTCTTGAATCAGAACCCGAGGGCGACGCTGCCCGCGAGGTCCAGGCGCT

[0496] GGCCGCTGAAATTAAATCAAAACTCATTTGAGTTAATGAGGTAAAGAGAAAAT

[0497] GAGCAAAAGCACAAACACGCTAAGTGCCGGCCGTCCGAGCGCACGCAGCAG

[0498] CAAGGCTGCAACGTTGGCCAGCCTGGCAGACACGCCAGCCATGAAGCGGGT

[0499] CAACTTTCAGTTGCCGGCGGAGGATCACACCAAGCTGAAGATGTACGCGGTA

[0500] CGCCAAGGCAAGACCATTACCGAGCTGCTATCTGAATACATCGCGCAGCTAC

[0501] CAGAGTAAATGAGCAAATGAATAAATGAGTAGATGAATTTTAGCGGCTAAAGG

[0502] AGGCGGCATGGAAAATCAAGAACAACCAGGCACCGACGCCGTGGAATGCCC

[0503] CATGTGTGGAGGAACGGGCGGTTGGCCAGGCGTAAGCGGCTGGGTTGCCT

[0504] GCCGGCCCTGCAATGGCACTGGAACCCCCAAGCCCGAGGAATCGGCGTGA

[0505] GCGGTCGCAAACCATCCGGCCCGGTACAAATCGGCGCGGCGCTGGGTGATG

[0506] ACCTGGTGGAGAAGTTGAAGGCCGCGCAGGCCGCCCAGCGGCAACGCATC

[0507] GAGGCAGAAGCACGCCCCGGTGAATCGTGGCAAGCGGCCGCTGATCGAATC

[0508] CGCAAAGAATCCCGGCAACCGCCGGCAGCCGGTGCGCCGTCGATTAGGAAG

[0509] CCGCCCAAGGGCGACGAGCAACCAGATTTTTTCGTTCCGATGCTCTATGACG

[0510] TGGGCACCCGCGATAGTCGCAGCATCATGGACGTGGCCGTTTTCCGTCTGTC

[0511] GAAGCGTGACCGACGAGCTGGCGAGGTGATCCGCTACGAGCTTCCAGACGG

[0512] GCACGTAGAGGTTTCCGCAGGGCCGGCCGGCATGGCCAGTGTGTGGGATTA

[0513] CGACCTGGTACTGATGGCGGTTTCCCATCTAACCGAATCCATGAACCGATAC

[0514] CGGGAAGGGAAGGGAGACAAGCCCGGCCGCGTGTTCCGTCCACACGTTGC

[0515] GGACGTACTCAAGTTCTGCCGGCGAGCCGATGGCGGAAAGCAGAAAGACGA

[0516] CCTGGTAGAAACCTGCATTCGGTTAAACACCACGCACGTTGCCATGCAGCGT

[0517] ACGAAGAAGGCCAAGAACGGCCGCCTGGTGACGGTATCCGAGGGTGAAGCC

[0518] TTGATTAGCCGCTACAAGATCGTAAAGAGCGAAACCGGGCGGCCGGAGTAC

[0519] ATCGAGATCGAGCTAGCTGATTGGATGTACCGCGAGATCACAGAAGGCAAGA

[0520] ACCCGGACGTGCTGACGGTTCACCCCGATTACTTTTTGATCGATCCCGGCAT

[0521] CGGCCGTTTTCTCTACCGCCTGGCACGCCGCGCCGCAGGCAAGGCAGAAGC

[0522] CAGATGGTTGTTCAAGACGATCTACGAACGCAGTGGCAGCGCCGGAGAGTT

[0523] CAAGAAGTTCTGTTTCACCGTGCGCAAGCTGATCGGGTCAAATGACCTGCCG

[0524] GAGTACGATTTGAAGGAGGAGGCGGGGCAGGCTGGCCCGATCCTAGTCATG CGCTACCGCAACCTGATCGAGGGCGAAGCATCCGCCGGTTCCTAATGTACG GAGCAGATGCTAGGGCAAATTGCCCTAGCAGGGGAAAAAGGTCGAAAAGGT CTCTTTCCTGTGGATAGCACGTACATTGGGAACCCAAAGCCGTACATTGGGA ACCGGAACCCGTACATTGGGAACCCAAAGCCGTACATTGGGAACCGGTCAC ACATGTAAGTGACTGATATAAAAGAGAAAAAAGGCGATTTTTCCGCCTAAAAC TCTTTAAAACTTATTAAAACTCTTAAAACCCGCCTGGCCTGTGCATAACTGTCT GGCCAGCGCACAGCCGAAGAGCTGCAAAAAGCGCCTACCCTTCGGTCGCTG CGCTCCCTACGCCCCGCCGCTTCGCGTCGGCCTATCGCGGCCGCTGGCCG CTCAAAAATGGCTGGCCTACGGCCAGGCAATCTACCAGGGCGCGGACAAGC CGCGCCGTCGCCACTCGACCGCCGGCGCCCACATCAAGGCACCCTGCCTCG CGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCGGAGAC GGTCACAGCTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGCCCGTCAGGG

[0525] CGCGTCAGCGGGTGTTGGCGGGTGTCGGGGCGCAGCCATGACCCAGTCAC GTAGCGATAGCGGAGTGTATACTGGCTTAACTATGCGGCATCAGAGCAGATT GTACTGAGAGTGCACCATATGCGGTGTGAAATACCGCACAGATGCGTAAGGA GAAAATACCGCATCAGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCG CTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAAT ACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAA GGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTC CATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGA GGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAA GCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTC CGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGG TATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAAC

[0526] CCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTC CAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAG

[0527] GATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGG CCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAA GCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACC ACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAA AAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAG TGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGCATTCTAGGTACTAAAA CAATTCATCCAGTAAAATATAATATTTTATTTTCTCCCAATCAGGCTTGATCCC CAGTAAGTCAAAAAATAGCTCGACATACTGTTCTTCCCCGATATCCTCCCTGA TCGACCGGACGCAGAAGGCAATGTCATACCACTTGTCCGCCCTGCCGCTTCT CCCAAGATCAATAAAGCCACTTACTTTGCCATCTTTCACAAAGATGTTGCTGT CTCCCAGGTCGCCGTGGGAAAAGACAAGTTCCTCTTCGGGCTTTTCCGTCTT TAAAAAATCATACAGCTCGCGCGGATCTTTAAATGGAGTGTCTTCTTCCCAGT TTTCGCAATCCACATCGGCCAGATCGTTATTCAGTAAGTAATCCAATTCGGCT

[0528] AAGCGGCTGTCTAAGCTATTCGTATAGGGACAATCCGATATGTCGATGGAGT

[0529] GAAAGAGCCTGATGCACTCCGCATACAGCTCGATAATCTTTTCAGGGCTTTGT

[0530] TCATCTTCATACTCTTCCGAGCAAAGGACGCCATCGGCCTCACTCATGAGCA

[0531] GATTGCTCCAGCCATCATGCCGTTCAAAGTGCAGGACCTTTGGAACAGGCAG

[0532] CTTTCCTTCCAGCCATAGCATCATGTCCTTTTCCCGTTCCACATCATAGGTGG

[0533] TCCCTTTATACCGGCTGTCCGTCATTTTTAAATATAGGTTTTCATTTTCTCCCA

[0534] CCAGCTTATATACCTTAGCAGGAGACATTCCTTCCGTATCTTTTACGCAGCGG

[0535] TATTTTTCGATCAGTTTTTTCAATTCCGGTGATATTCTCATTTTAGCCATTTATT

[0536] ATTTCCTTCCTCTTTTCTACAGTATTTAAAGATACCCCAAGAAGCTAATTATAA

[0537] CAAGACGAACTCCAATTCACTGTTCCTTGCATTCTAAAACCTTAAATACCAGA

[0538] AAACAGCTTTTTCAAAGTTGTTTTCAAAGTTGGCGTATAACATAGTATCGACG

[0539] GAGCCGATTTTGAAACCGCGGTGATCACAGGCAGCAACGCTCTGTCATCGTT

[0540] ACAATCAACATGCTACCCTCCGCGAGATCATCCGTGTTTCAAACCCGGCAGC

[0541] TTAGTTGCCGTTCTTCCGAATAGCATCGGTAACATGAGCAAAGTCTGCCGCCT

[0542] TACAACGGCTCTCCCGCTGACGCCGTCCCGGACTGATGGGCTGCCTGTATC

[0543] GAGTGGTGATTTTGTGCCGAGCTGCCGGTCGGGGAGCTGTTGGCTGGCTGG

[0544] TGGCAGGATATATTGTGGTGTAAACAAATTGACGCTTAGACAACTTAATAACA

[0545] CATTGCGGACGTTTTTAATGTACTGAATTAACGCCGAATTGAATTCGAGCTCG

[0546] GTACCGGGGGATCTGGATTTTAGTACTGGATTTTGGTTTTAGGAATTAGAAAT

[0547] >

[0548] TTTATTGATAGAAGTATTTTACAAATACAAATACATACTAAGGGTTTCTTATATG

[0549] CTCAACACATGAGCGAAACCCTATAGGAACCCTAATTCCCTTATCTGGGAACT

[0550] ACTCACACATTATTATGGAGAAACTCGAGCTTGTCGATCGACTCAAATCTCGG

[0551] TGACGGGCAGGACCGGACGGGGCGGTACCGGCAGGCTGAAGTCCAGCTGC

[0552] CAGAAACCCACGTCATGCCAGTTCCCGTGCTTGAAGCCGGCCGCCCGCAGC

[0553] ATGCCGCGGGGGGCATATCCGAGCGCCTCGTGCATGCGCACGCTCGGGTC

[0554] GTTGGGCAGCCCGATGACAGCGACCACGCTCTTGAAGCCCTGTGCCTCCAG

[0555] GGACTTCAGCAGGTGGGTGTAGAGCGTGGAGCCCAGTCCCGTCCGCTGGTG

[0556] GCGGGGGGAGACGTACACGGTCGACTCGGCCGTCCAGTCGTAGGCGTTGC

[0557] GTGCCTTCCAGGGGCCCGCGTAGGCGATGCCGGCGACCTCGCCGTCCACCT

[0558] CGGCGACGAGCCAGGGATAGCGCTCCCGCAGACGGACGAGGTCGTCCGTC

[0559] CACTCCTGCGGTTCCTGCGGCTCGGTACGGAAGTTGACCGTGCTTGTCTCGA

[0560] TGTAGTGGTTGACGATGGTGCAGACCGCCGGCATGTCCGCCTCGGTGGCAC

[0561] GGCGGATGTCGGCCGGGCGTCGTTCTGGGCTCATGGTAGACGATCTGCGAA

[0562] AGCTCGAGAGAGATAGATTTGTAGAGAGAGACTGGTGATTTCAGCGTGTCCT

[0563] CTCCAAATGAAATGAACTTCCTTATATAGAGGAAGGGTCTTGCGAAGGATAGT

[0564] GGGATTGTGCGTCATCCCTTACGTCAGTGGAGATATCACATCAATCCACTTGC

[0565] TTTGAAGACGTGGTTGGAACGTCTTCTTTTTCCACGATGCTCCTCGTGGGTG GGGGTCCATCTTTGGGACCACTGTCGGCAGAGGCATCTTGAACGATAGCCTT TCCTTTATCGCAATGATGGCATTTGTAGGTGCCACCTTCCTTTTCTACTGTCCT TTTGATGAAGTGACAGATAGCTGGGCAATGGAATCCGAGGAGGTTTCCCGAT ATTACCCTTTGTTGAAAAGTCTCAATAGCCCTTTGGTCTTCTGAGACTGTATCT

[0566] TTGATATTCTTGGAGTAGACGAGAGTGTCGTGCTCCACCATGTTCACATCAAT

[0567] CCACTTGCTTTGAAGACGTGGTTGGAACGTCTTCTTTTTCCACGATGCTCCTC

[0568] GTGGGTGGGGGTCCATCTTTGGGACCACTGTCGGCAGAGGCATCTTGAACG ATAGCCTTTCCTTTATCGCAATGATGGCATTTGTAGGTGCCACCTTCCTTTTCT

[0569] ACTGTCCTTTTGATGAAGTGACAGATAGCTGGGCAATGGAATCCGAGGAGGT TTCCCGATATTACCCTTTGTTGAAAAGTCTCAATAGCCCTTTGGTCTTCTGAGA CTGTATCTTTGATATTCTTGGAGTAGACGAGAGTGTCGTGCTCCACCATGTTG GCGTACCCGGGGATCCTCTAGAGTCGACCTGCAGGCATGCAAGCT

[0570] Bovine Collagen alpha-1 (I) Expression vector 8;

[0571] SEQ ID NO: 12:

[0572] AGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCG

[0573] CCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAA

[0574] CCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTG

[0575] CGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCC

[0576] CTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTC

[0577] GGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCA

[0578] GCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTA

[0579] AGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAG

[0580] CGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGG

[0581] CTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCT

[0582] TCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAG

[0583] CGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTC

[0584] AAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAAC

[0585] TCACGTTAAGGGATTTTGGTCATGCATTCTAGGTACTAAAACAATTCATCCAG

[0586] TAAAATATAATATTTTATTTTCTCCCAATCAGGCTTGATCCCCAGTAAGTCAAA

[0587] AAATAGCTCGACATACTGTTCTTCCCCGATATCCTCCCTGATCGACCGGACG

[0588] CAGAAGGCAATGTCATACCACTTGTCCGCCCTGCCGCTTCTCCCAAGATCAA

[0589] TAAAGCCACTTACTTTGCCATCTTTCACAAAGATGTTGCTGTCTCCCAGGTCG

[0590] CCGTGGGAAAAGACAAGTTCCTCTTCGGGCTTTTCCGTCTTTAAAAAATCATA

[0591] CAGCTCGCGCGGATCTTTAAATGGAGTGTCTTCTTCCCAGTTTTCGCAATCCA

[0592] CATCGGCCAGATCGTTATTCAGTAAGTAATCCAATTCGGCTAAGCGGCTGTCT AAGCTATTCGTATAGGGACAATCCGATATGTCGATGGAGTGAAAGAGCCTGA TGCACTCCGCATACAGCTCGATAATCTTTTCAGGGCTTTGTTCATCTTCATACT CTTCCGAGCAAAGGACGCCATCGGCCTCACTCATGAGCAGATTGCTCCAGCC

[0593] ATCATGCCGTTCAAAGTGCAGGACCTTTGGAACAGGCAGCTTTCCTTCCAGC

[0594] CATAGCATCATGTCCTTTTCCCGTTCCACATCATAGGTGGTCCCTTTATACCG

[0595] GCTGTCCGTCATTTTTAAATATAGGTTTTCATTTTCTCCCACCAGCTTATATAC

[0596] CTTAGCAGGAGACATTCCTTCCGTATCTTTTACGCAGCGGTATTTTTCGATCA

[0597] GTTTTTTCAATTCCGGTGATATTCTCATTTTAGCCATTTATTATTTCCTTCCTCT

[0598] TTTCTACAGTATTTAAAGATACCCCAAGAAGCTAATTATAACAAGACGAACTCC

[0599] AATTCACTGTTCCTTGCATTCTAAAACCTTAAATACCAGAAAACAGCTTTTTCA

[0600] AAGTTGTTTTCAAAGTTGGCGTATAACATAGTATCGACGGAGCCGATTTTGAA

[0601] ACCGCGGTGATCACAGGCAGCAACGCTCTGTCATCGTTACAATCAACATGCT

[0602] ACCCTCCGCGAGATCATCCGTGTTTCAAACCCGGCAGCTTAGTTGCCGTTCT

[0603] TCCGAATAGCATCGGTAACATGAGCAAAGTCTGCCGCCTTACAACGGCTCTC

[0604] CCGCTGACGCCGTCCCGGACTGATGGGCTGCCTGTATCGAGTGGTGATTTT

[0605] GTGCCGAGCTGCCGGTCGGGGAGCTGTTGGCTGGCTGGTGGCAGGATATAT

[0606] TGTGGTGTAAACAAATTGACGCTTAGACAACTTAATAACACATTGCGGACGTT

[0607] >

[0608] TTTAATGTACTGAATTAACGCCGAATTGAATTCGAGCTCGGTGGAGGATCTGG

[0609] ATTTTAGTACTGGATTTTGGTTTTAGGAATTAGAAATTTTATTGATAGAAGTATT

[0610] TTACAAATACAAATACATACTAAGGGTTTCTTATATGCTCAACACATGAGCGAA

[0611] ACCCTATAGGAACCCTAATTCCCTTATCTGGGAACTACTCACACATTATTATG

[0612] GAGAAACTATTCACTCTCCTTCTCAAACGTTTCTGTTCCCTCTGGACAGCCCC

[0613] GACTGACGTTGGACTCAACCTCAATGGGCGCTAATTGTAATAGCTCCTGTTTG

[0614] CTTCTTATGGCCATTTCGCAAAGGTGTTGCAGTGTTCTGCTACCACCACTCAC

[0615] TGTAATTGACACACCTCTGAAACGGATGGAGTAAGTGCAACCAATTTGATCAA

[0616] ATCCAAAGAAACGACTAGCAGCGTAATTTACTGAGTCACCAGTCCAGGAACC

[0617] TAGAACTCGATGGAGAGAGGCCTCTGTCCGATCATAGCGTAGGTAGCGTTTG

[0618] AAAACTACCTTGCCAAATCCCCAGGACTCTTTAAATCCCAGTGGGTTGTCCTG

[0619] GTTGGAGTTTGTTTCGTCATTGTGTAAACGCCATTCTGTCCAAGATGGGCTCT

[0620] CATCGGGTAACTTAAATGGGCTGGTAGTGCCGCCAGAACCACCGTCCCATCT

[0621] CTCTGAATTGGCCTGCTCTCTTGCATCATTACCCTGGATTGCTCTCTCCATTG

[0622] TCCTCTCCAAATGAAATGAACTTCCTTATATAGAGGAAGGGTCTTGCGAAGGA

[0623] TAGTGGGATTGTGCGTCATCCCTTACGTCAGTGGAGATGTCACATCAATCCA

[0624] CTTGCTTTGTAGACGTGGTTGGAACCTCTTCTTTTTCCACGATGCTCCTCGTG

[0625] GGTGGGGGTCCATCTTTGGGACCACTGTCGGCAGAGAGATCTTGAATGATAG

[0626] CCTTTCCTTTATCGCAATGATGGCATTTGTAGGAGCCACCTTCCTTTTCTACT

[0627] GTCCTTTCGATGAAGTGACAGATAGCTGGGCAATGGAATCCGAGGAGGTTTC

[0628] CCGAAATTATCCTTTGTTGAAAAGTCTCAAACAATAGGCAACCGTGGACTTCT

[0629] TCACTAGCTCATCGAGATAGCAATGCCACTAGCTAATTTCTTACGTTGTATCTT

[0630] ATTTGTTTTACTTTGGGGCATGACATGGTTAAACCCCATCAAAGAGAAAGTGT TTCATCCACTAGTCAATATACGAATTCATTCGAATTTATCCCTGTAAATCCTAG TTCTTAGGATGAACTGGTGTAATAAACAGCAAAAAAAAATAAAAAAAAATAAAT CTGGAATCATTCGACCACCTCAATAAACTAAAGCTACCAATTACCACAATATA GTC ATC C ATATC C ACTTAGATATAAAAGATAAAAGTAAAC AAATATTAAATTTC ATATGCACGCATAGGAAACTCATGATCTTATCTTTTAAATAGACATCTAGTTTT CTTAGGTTATAAATAGACATTTTGTCCTAGAACTTCTTCACTACTAAAACCTAG CCTCAAAAAGCAGGCTGCCACCATGCATCACCATCATCATCATGGATCTGAG AATTTATATTTTCAAGGTCAATTGAGTTATGGATACGACGAAAAATCAACTGGA ATATCAGTGCCAGGACCAATGGGTCCATCAGGACCTAGGGGTCTTCCAGGG CCACCTGGCGCTCCTGGTCCTCAGGGATTTCAAGGTCCTCCAGGTGAACCTG GAGAGCCTGGTGCTTCTGGTCCAATGGGACCAAGAGGCCCTCCTGGTCCAC CTGGTAAAAATGGTGATGATGGTGAGGCTGGAAAGCCTGGTCGTCCTGGAG AAAGAGGACCACCAGGACCACAGGGAGCTAGAGGTTTGCCAGGTACAGCCG GTCTTCCAGGAATGAAAGGACACAGGGGTTTTTCTGGTCTTGATGGTGCTAA GGGAGACGCAGGACCTGCTGGACCAAAGGGTGAACCTGGATCTCCTGGTGA AAATGGTGCTCCTGGACAAATGGGACCTAGAGGATTGCCTGGCGAAAGAGG AAGACCTGGTGCTCCTGGACCAGCTGGAGCTAGAGGAAATGATGGAGCTAC TGGTGCTGCCGGACCACCAGGACCAACTGGACCAGCTGGACCACCTGGTTT TCCAGGAGCAGTTGGGGCTAAGGGAGAAGGTGGACCACAAGGACCTAGAGG ATCAGAAGGACCTCAAGGAGTTAGGGGAGAACCAGGTCCTCCTGGTCCTGC TGGAGCTGCTGGACCCGCTGGTAACCCAGGAGCAGATGGACAACCTGGAGC AAAAGGGGCTAATGGTGCTCCTGGTATTGCAGGTGCTCCTGGATTCCCTGGT GCTAGAGGACCTTCAGGACCTCAGGGACCATCCGGTCCTCCAGGACCAAAG GGAAACTCTGGTGAACCAGGGGCTCCTGGATCTAAGGGAGATACTGGAGCT AAGGGTGAGCCAGGACCTACAGGAATTCAAGGACCACCAGGACCTGCAGGA GAGGAAGGTAAGAGGGGTGCTAGAGGTGAACCTGGACCAGCTGGATTGCCA GGCCCACCTGGAGAGAGGGGTGGTCCAGGTTCTAGAGGTTTTCCAGGAGCT GATGGAGTTGCTGGTCCAAAGGGTCCTGCTGGTGAGAGAGGTGCACCTGGA CCAGCAGGACCTAAAGGATCACCTGGAGAAGCTGGTCGTCCTGGTGAGGCT GGTCTTCCAGGTGCTAAGGGCCTTACAGGATCTCCAGGATCACCAGGTCCAG ATGGAAAGACTGGACCACCAGGTCCTGCAGGACAAGATGGAAGGCCTGGTC CACCTGGACCACCTGGTGCTAGGGGACAAGCTGGTGTTATGGGATTCCCAG GTCCAAAGGGTGCAGCTGGTGAGCCTGGTAAAGCTGGTGAAAGAGGAGTTC CAGGACCTCCTGGTGCTGTTGGACCTGCTGGTAAAGATGGTGAAGCAGGTG CTCAGGGTCCACCAGGTCCAGCCGGTCCAGCTGGTGAGAGGGGAGAGCAA GGTCCTGCTGGATCTCCTGGATTTCAAGGTTTGCCTGGACCTGCTGGACCTC CAGGAGAGGCTGGTAAACCTGGTGAGCAGGGTGTGCCTGGTGATCTTGGTG CTCCTGGACCTAGTGGCGCAAGAGGCGAAAGGGGATTTCCAGGTGAAAGAG GAGTTCAGGGACCACCAGGTCCAGCTGGTCCAAGAGGCGCTAATGGAGCTC

[0631] CAGGAAATGATGGAGCAAAGGGTGATGCTGGCGCTCCAGGAGCTCCTGGTT

[0632] CTCAAGGTGCACCTGGATTGCAAGGAATGCCAGGAGAGAGAGGTGCAGCTG

[0633] GTTTGCCGGGACCAAAGGGTGATAGAGGTGATGCTGGTCCTAAGGGAGCTG

[0634] ATGGAGCTCCAGGAAAGGATGGTGTGAGAGGATTGACTGGTCCAATTGGAC

[0635] CACCAGGTCCTGCTGGAGCACCAGGAGATAAAGGAGAAGCTGGTCCTTCAG

[0636] GACCAGCCGGTCCAACTGGAGCTAGAGGAGCTCCAGGAGATAGAGGTGAAC

[0637] CAGGACCACCAGGACCTGCTGGGTTTGCAGGTCCACCTGGTGCTGATGGAC

[0638] AACCTGGTGCTAAAGGAGAACCAGGCGATGCTGGTGCTAAGGGAGATGCTG

[0639] GCCCACCTGGTCCAGCTGGTCCTGCTGGGCCACCAGGTCCTATCGGAAATG

[0640] TTGGTGCTCCAGGGCCTAAAGGAGCTAGAGGATCAGCAGGTCCCCCAGGTG

[0641] CTACAGGTTTTCCAGGGGCTGCTGGAAGAGTGGGTCCCCCAGGTCCTTCTG

[0642] GGAATGCAGGACCACCAGGTCCTCCAGGACCAGCTGGAAAAGAAGGAAGTA

[0643] AGGGACCAAGGGGAGAAACTGGACCTGCAGGACGTCCAGGTGAAGTTGGTC

[0644] CACCTGGACCTCCAGGTCCAGCTGGAGAGAAAGGAGCTCCAGGAGCTGATG

[0645] GTCCAGCAGGTGCTCCTGGTACTCCAGGTCCACAAGGTATTGCTGGTCAAAG

[0646] GGGAGTGGTTGGTCTCCCAGGTCAGAGAGGTGAAAGAGGTTTCCCAGGTTT

[0647] GCCTGGACCATCTGGAGAACCAGGTAAGCAGGGCCCATCTGGAGCTTCTGG

[0648] TGAGAGAGGGCCACCTGGACCTATGGGACCTCCTGGGCTTGCTGGACCTCC

[0649] AGGGGAATCTGGTAGGGAGGGTGCTCCAGGTGCTGAAGGATCACCTGGTAG

[0650] AGATGGATCACCTGGAGCTAAAGGTGATAGAGGGGAAACAGGTCCAGCAGG

[0651] TCCACCTGGCGCTCCTGGTGCTCCAGGAGCACCAGGGCCAGTTGGACCTGC

[0652] TGGAAAGTCTGGAGATAGGGGAGAGACTGGACCAGCTGGACCTGCTGGACC

[0653] AATCGGTCCTGTTGGCGCTAGAGGTCCTGCTGGACCACAAGGTCCTAGGGG

[0654] AGATAAAGGTGAAACAGGTGAACAAGGAGATAGAGGAATTAAAGGTCATAGA

[0655] GGGTTTTCTGGTTTGCAAGGACCTCCAGGACCTCCTGGATCTCCAGGAGAGC

[0656] AAGGACCATCTGGTGCAAGCGGACCTGCTGGTCCTAGAGGACCTCCAGGTT

[0657] CAGCTGGCTCACCTGGTAAGGATGGATTAAATGGTTTGCCTGGACCAATAGG

[0658] TCCACCAGGACCTAGAGGAAGGACAGGTGATGCAGGACCAGCTGGTCCACC

[0659] TGGTCCACCAGGACCACCAGGACCACCAGGTCCACCTTCTGGTGGATATGAT

[0660] CTTAGTTTTCTTCCTCAACCTCCCCAGGAGAAAGCTCATGATGGTGGTAGATA

[0661] TTATAGGGCTTAAACCCAGCTTTCTTGTACAAAGTGGTAGCTCGAATTTCCCC

[0662] GATCGTTCAAACATTTGGCAATAAAGTTTCTTAAGATTGAATCCTGTTGCCGG

[0663] TCTTGCGATGATTATCATATAATTTCTGTTGAATTACGTTAAGCATGTAATAATT

[0664] AACATGTAATGCATGACGTTATTTATGAGATGGGTTTTTATGATTAGAGTCCC

[0665] GCAATTATACATTTAATACGCGATAGAAAACAAAATATAGCGCGCAAACTAGG

[0666] ATAAATTATCGCGCGCGGTGTCATCTATGTTACTAGATCGGGAAAGCTTTTAA

[0667] ACTATCAGTGTTTGACAGGATATATTGGCGGGTAAACCTAAGAGAAAAGAGC GTTTATTAGAATAATCGGATATTTAAAAGGGCGTGAAAAGGTTTATCCGTTCG

[0668] TCCATTTGTATGTGCATGCCAACCACAGGGTTCCCCTCGGGATCAAAGTACTT

[0669] TGATCCAACCCCTCCGCTGCTATAGTGCAGTCGGCTTCTGACGTTCAGTGCA

[0670] GCCGTCTTCTGAAAACGACATGTCGCACAAGTCCTAAGTTACGCGACAGGCT

[0671] GCCGCCCTGCCCTTTTCCTGGCGTTTTCTTGTCGCGTGTTTTAGTCGCATAAA

[0672] GTAGAATACTTGCGACTAGAACCGGAGACATTACGCCATGAACAAGAGCGCC

[0673] GCCGCTGGCCTGCTGGGCTATGCCCGCGTCAGCACCGACGACCAGGACTTG

[0674] ACCAACCAACGGGCCGAACTGCACGCGGCCGGCTGCACCAAGCTGTTTTCC

[0675] GAGAAGATCACCGGCACCAGGCGCGACCGCCCGGAGCTGGCCAGGATGCT

[0676] TGACCACCTACGCCCTGGCGACGTTGTGACAGTGACCAGGCTAGACCGCCT

[0677] GGCCCGCAGCACCCGCGACCTACTGGACATTGCCGAGCGCATCCAGGAGG

[0678] CCGGCGCGGGCCTGCGTAGCCTGGCAGAGCCGTGGGCCGACACCACCACG

[0679] CCGGCCGGCCGCATGGTGTTGACCGTGTTCGCCGGCATTGCCGAGTTCGAG

[0680] CGTTCCCTAATCATCGACCGCACCCGGAGCGGGCGCGAGGCCGCCAAGGC

[0681] CCGAGGCGTGAAGTTTGGCCCCCGCCCTACCCTCACCCCGGCACAGATCGC

[0682] GCACGCCCGCGAGCTGATCGACCAGGAAGGCCGCACCGTGAAAGAGGCGG

[0683] CTGCACTGCTTGGCGTGCATCGCTCGACCCTGTACCGCGCACTTGAGCGCA

[0684] GCGAGGAAGTGACGCCCACCGAGGCCAGGCGGCGCGGTGCCTTCCGTGAG

[0685] GACGCATTGACCGAGGCCGACGCCCTGGCGGCCGCCGAGAATGAACGCCA

[0686] AGAGGAACAAGCATGAAACCGCACCAGGACGGCCAGGACGAACCGTTTTTC

[0687] ATTACCGAAGAGATCGAGGCGGAGATGATCGCGGCCGGGTACGTGTTCGAG

[0688] CCGCCCGCGCACGTCTCAACCGTGCGGCTGCATGAAATCCTGGCCGGTTTG

[0689] TCTGATGCCAAGCTGGCGGCCTGGCCGGCCAGCTTGGCCGCTGAAGAAACC

[0690] GAGCGCCGCCGTCTAAAAAGGTGATGTGTATTTGAGTAAAACAGCTTGCGTC

[0691] ATGCGGTCGCTGCGTATATGATGCGATGAGTAAATAAACAAATACGCAAGGG

[0692] GAACGCATGAAGGTTATCGCTGTACTTAACCAGAAAGGCGGGTCAGGCAAGA

[0693] CGACCATCGCAACCCATCTAGCCCGCGCCCTGCAACTCGCCGGGGCCGATG

[0694] TTCTGTTAGTCGATTCCGATCCCCAGGGCAGTGCCCGCGATTGGGCGGCCG

[0695] TGCGGGAAGATCAACCGCTAACCGTTGTCGGCATCGACCGCCCGACGATTG

[0696] ACCGCGACGTGAAGGCCATCGGCCGGCGCGACTTCGTAGTGATCGACGGAG

[0697] CGCCCCAGGCGGCGGACTTGGCTGTGTCCGCGATCAAGGCAGCCGACTTCG

[0698] TGCTGATTCCGGTGCAGCCAAGCCCTTACGACATATGGGCCACCGCCGACCT

[0699] GGTGGAGCTGGTTAAGCAGCGCATTGAGGTCACGGATGGAAGGCTACAAGC

[0700] GGCCTTTGTCGTGTCGCGGGCGATCAAAGGCACGCGCATCGGCGGTGAGGT

[0701] TGCCGAGGCGCTGGCCGGGTACGAGCTGCCCATTCTTGAGTCCCGTATCAC

[0702] GCAGCGCGTGAGCTACCCAGGCACTGCCGCCGCCGGCACAACCGTTCTTGA

[0703] ATCAGAACCCGAGGGCGACGCTGCCCGCGAGGTCCAGGCGCTGGCCGCTG

[0704] AAATTAAATCAAAACTCATTTGAGTTAATGAGGTAAAGAGAAAATGAGCAAAA GCACAAACACGCTAAGTGCCGGCCGTCCGAGCGCACGCAGCAGCAAGGCTG

[0705] CAACGTTGGCCAGCCTGGCAGACACGCCAGCCATGAAGCGGGTCAACTTTC

[0706] AGTTGCCGGCGGAGGATCACACCAAGCTGAAGATGTACGCGGTACGCCAAG

[0707] GCAAGACCATTACCGAGCTGCTATCTGAATACATCGCGCAGCTACCAGAGTA

[0708] AATGAGCAAATGAATAAATGAGTAGATGAATTTTAGCGGCTAAAGGAGGCGG

[0709] CATGGAAAATCAAGAACAACCAGGCACCGACGCCGTGGAATGCCCCATGTGT

[0710] GGAGGAACGGGCGGTTGGCCAGGCGTAAGCGGCTGGGTTGCCTGCCGGCC

[0711] CTGCAATGGCACTGGAACCCCCAAGCCCGAGGAATCGGCGTGAGCGGTCGC

[0712] AAACCATCCGGCCCGGTACAAATCGGCGCGGCGCTGGGTGATGACCTGGTG

[0713] GAGAAGTTGAAGGCCGCGCAGGCCGCCCAGCGGCAACGCATCGAGGCAGA

[0714] AGCACGCCCCGGTGAATCGTGGCAAGCGGCCGCTGATCGAATCCGCAAAGA

[0715] ATCCCGGCAACCGCCGGCAGCCGGTGCGCCGTCGATTAGGAAGCCGCCCA

[0716] AGGGCGACGAGCAACCAGATTTTTTCGTTCCGATGCTCTATGACGTGGGCAC

[0717] CCGCGATAGTCGCAGCATCATGGACGTGGCCGTTTTCCGTCTGTCGAAGCGT

[0718] GACCGACGAGCTGGCGAGGTGATCCGCTACGAGCTTCCAGACGGGCACGTA

[0719] GAGGTTTCCGCAGGGCCGGCCGGCATGGCCAGTGTGTGGGATTACGACCTG

[0720] GTACTGATGGCGGTTTCCCATCTAACCGAATCCATGAACCGATACCGGGAAG

[0721] GGAAGGGAGACAAGCCCGGCCGCGTGTTCCGTCCACACGTTGCGGACGTAC

[0722] TCAAGTTCTGCCGGCGAGCCGATGGCGGAAAGCAGAAAGACGACCTGGTAG

[0723] AAACCTGCATTCGGTTAAACACCACGCACGTTGCCATGCAGCGTACGAAGAA

[0724] GGCCAAGAACGGCCGCCTGGTGACGGTATCCGAGGGTGAAGCCTTGATTAG

[0725] CCGCTACAAGATCGTAAAGAGCGAAACCGGGCGGCCGGAGTACATCGAGAT

[0726] CGAGCTAGCTGATTGGATGTACCGCGAGATCACAGAAGGCAAGAACCCGGA

[0727] CGTGCTGACGGTTCACCCCGATTACTTTTTGATCGATCCCGGCATCGGCCGT

[0728] TTTCTCTACCGCCTGGCACGCCGCGCCGCAGGCAAGGCAGAAGCCAGATGG

[0729] TTGTTCAAGACGATCTACGAACGCAGTGGCAGCGCCGGAGAGTTCAAGAAGT

[0730] TCTGTTTCACCGTGCGCAAGCTGATCGGGTCAAATGACCTGCCGGAGTACGA

[0731] TTTGAAGGAGGAGGCGGGGCAGGCTGGCCCGATCCTAGTCATGCGCTACCG

[0732] CAACCTGATCGAGGGCGAAGCATCCGCCGGTTCCTAATGTACGGAGCAGAT

[0733] GCTAGGGCAAATTGCCCTAGCAGGGGAAAAAGGTCGAAAAGGTCTCTTTCCT

[0734] GTGGATAGCACGTACATTGGGAACCCAAAGCCGTACATTGGGAACCGGAAC

[0735] CCGTACATTGGGAACCCAAAGCCGTACATTGGGAACCGGTCACACATGTAAG

[0736] TGACTGATATAAAAGAGAAAAAAGGCGATTTTTCCGCCTAAAACTCTTTAAAA

[0737] CTTATTAAAACTCTTAAAACCCGCCTGGCCTGTGCATAACTGTCTGGCCAGCG

[0738] CACAGCCGAAGAGCTGCAAAAAGCGCCTACCCTTCGGTCGCTGCGCTCCCT

[0739] ACGCCCCGCCGCTTCGCGTCGGCCTATCGCGGCCGCTGGCCGCTCAAAAAT

[0740] GGCTGGCCTACGGCCAGGCAATCTACCAGGGCGCGGACAAGCCGCGCCGT

[0741] CGCCACTCGACCGCCGGCGCCCACATCAAGGCACCCTGCCTCGCGCGTTTC GGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCGGAGACGGTCACA GCTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGCCCGTCAGGGCGCGTCA GCGGGTGTTGGCGGGTGTCGGGGCGCAGCCATGACCCAGTCACGTAGCGA TAGCGGAGTGTATACTGGCTTAACTATGCGGCATCAGAGCAGATTGTACTGA

[0742] GAGTGCACCATATGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATA CCGCATCAGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTC GTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTAT CCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGC

[0743] AAA

[0744] Bovine Collagen alpha-1 (I) Expression vector 9;

[0745] SEQ ID NO: 15:

[0746] AGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCG

[0747] CCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAA CCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTG CGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCC CTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTC

[0748] GGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCA GCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTA AGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAG CGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGG

[0749] CTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCT TCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAG CGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTC AAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAAC

[0750] TCACGTTAAGGGATTTTGGTCATGCATTCTAGGTACTAAAACAATTCATCCAG TAAAATATAATATTTTATTTTCTCCCAATCAGGCTTGATCCCCAGTAAGTCAAA AAATAGCTCGACATACTGTTCTTCCCCGATATCCTCCCTGATCGACCGGACG CAGAAGGCAATGTCATACCACTTGTCCGCCCTGCCGCTTCTCCCAAGATCAA

[0751] TAAAGCCACTTACTTTGCCATCTTTCACAAAGATGTTGCTGTCTCCCAGGTCG CCGTGGGAAAAGACAAGTTCCTCTTCGGGCTTTTCCGTCTTTAAAAAATCATA CAGCTCGCGCGGATCTTTAAATGGAGTGTCTTCTTCCCAGTTTTCGCAATCCA CATCGGCCAGATCGTTATTCAGTAAGTAATCCAATTCGGCTAAGCGGCTGTCT

[0752] AAGCTATTCGTATAGGGACAATCCGATATGTCGATGGAGTGAAAGAGCCTGA TGCACTCCGCATACAGCTCGATAATCTTTTCAGGGCTTTGTTCATCTTCATACT CTTCCGAGCAAAGGACGCCATCGGCCTCACTCATGAGCAGATTGCTCCAGCC ATCATGCCGTTCAAAGTGCAGGACCTTTGGAACAGGCAGCTTTCCTTCCAGC

[0753] CATAGCATCATGTCCTTTTCCCGTTCCACATCATAGGTGGTCCCTTTATACCG GCTGTCCGTCATTTTTAAATATAGGTTTTCATTTTCTCCCACCAGCTTATATAC

[0754] CTTAGCAGGAGACATTCCTTCCGTATCTTTTACGCAGCGGTATTTTTCGATCA

[0755] GTTTTTTCAATTCCGGTGATATTCTCATTTTAGCCATTTATTATTTCCTTCCTCT

[0756] TTTCTACAGTATTTAAAGATACCCCAAGAAGCTAATTATAACAAGACGAACTCC

[0757] AATTCACTGTTCCTTGCATTCTAAAACCTTAAATACCAGAAAACAGCTTTTTCA

[0758] AAGTTGTTTTCAAAGTTGGCGTATAACATAGTATCGACGGAGCCGATTTTGAA

[0759] ACCGCGGTGATCACAGGCAGCAACGCTCTGTCATCGTTACAATCAACATGCT

[0760] ACCCTCCGCGAGATCATCCGTGTTTCAAACCCGGCAGCTTAGTTGCCGTTCT

[0761] TCCGAATAGCATCGGTAACATGAGCAAAGTCTGCCGCCTTACAACGGCTCTC

[0762] CCGCTGACGCCGTCCCGGACTGATGGGCTGCCTGTATCGAGTGGTGATTTT

[0763] GTGCCGAGCTGCCGGTCGGGGAGCTGTTGGCTGGCTGGTGGCAGGATATAT

[0764] TGTGGTGTAAACAAATTGACGCTTAGACAACTTAATAACACATTGCGGACGTT

[0765] TTTAATGTACTGAATTAACGCCGAATTGAATTCGAGCTCGGTGGAGGATCTGG

[0766] ATTTTAGTACTGGATTTTGGTTTTAGGAATTAGAAATTTTATTGATAGAAGTATT

[0767] TTACAAATACAAATACATACTAAGGGTTTCTTATATGCTCAACACATGAGCGAA

[0768] ACCCTATAGGAACCCTAATTCCCTTATCTGGGAACTACTCACACATTATTATG

[0769] GAGAAACTATTCACTCTCCTTCTCAAACGTTTCTGTTCCCTCTGGACAGCCCC

[0770] GACTGACGTTGGACTCAACCTCAATGGGCGCTAATTGTAATAGCTCCTGTTTG

[0771] CTTCTTATGGCCATTTCGCAAAGGTGTTGCAGTGTTCTGCTACCACCACTCAC

[0772] TGTAATTGACACACCTCTGAAACGGATGGAGTAAGTGCAACCAATTTGATCAA

[0773] ATCCAAAGAAACGACTAGCAGCGTAATTTACTGAGTCACCAGTCCAGGAACC

[0774] TAGAACTCGATGGAGAGAGGCCTCTGTCCGATCATAGCGTAGGTAGCGTTTG

[0775] AAAACTACCTTGCCAAATCCCCAGGACTCTTTAAATCCCAGTGGGTTGTCCTG

[0776] GTTGGAGTTTGTTTCGTCATTGTGTAAACGCCATTCTGTCCAAGATGGGCTCT

[0777] CATCGGGTAACTTAAATGGGCTGGTAGTGCCGCCAGAACCACCGTCCCATCT

[0778] CTCTGAATTGGCCTGCTCTCTTGCATCATTACCCTGGATTGCTCTCTCCATTG

[0779] TCCTCTCCAAATGAAATGAACTTCCTTATATAGAGGAAGGGTCTTGCGAAGGA

[0780] TAGTGGGATTGTGCGTCATCCCTTACGTCAGTGGAGATGTCACATCAATCCA

[0781] CTTGCTTTGTAGACGTGGTTGGAACCTCTTCTTTTTCCACGATGCTCCTCGTG

[0782] GGTGGGGGTCCATCTTTGGGACCACTGTCGGCAGAGAGATCTTGAATGATAG

[0783] CCTTTCCTTTATCGCAATGATGGCATTTGTAGGAGCCACCTTCCTTTTCTACT

[0784] GTCCTTTCGATGAAGTGACAGATAGCTGGGCAATGGAATCCGAGGAGGTTTC

[0785] CCGAAATTATCCTTTGTTGAAAAGTCTCAAACAATAGGCAACCGTGGACTTCT

[0786] TCACTAGCTCATCGAGATAGCAATGCCACTAGCTAATTTCTTACGTTGTATCTT

[0787] ATTTGTTTTACTTTGGGGCATGACATGGTTAAACCCCATCAAAGAGAAAGTGT

[0788] TTCATCCACTAGTCAATATACGAATTCATTCGAATTTATCCCTGTAAATCCTAG

[0789] TTCTTAGGATGAACTGGTGTAATAAACAGCAAAAAAAAATAAAAAAAAATAAAT

[0790] CTGGAATCATTCGACCACCTCAATAAACTAAAGCTACCAATTACCACAATATA GTC ATC C ATATC C ACTTAGATATAAAAGATAAAAGTAAAC AAATATTAAATTTC ATATGCACGCATAGGAAACTCATGATCTTATCTTTTAAATAGACATCTAGTTTT CTTAGGTTATAAATAGACATTTTGTCCTAGAACTTCTTCACTACTAAAACCTAG CCTCAAAAAGCAGGCTGCCACCATGGTGAGCAGTGGTGAGGACATCTTTTCA GGCCTAGTCCCGATACTTATTGAACTTGAAGGAGACGTCAATGGGCACAGAT TCTCAGTACGAGGCGAGGGATACGGGGATGCTTCCAATGGAAAGCTTGAGAT AAAATTTATATGTACAACTGGAAGGCTTCCTGTCCCATGGCCAACTTTAGTTA CCACTCTTAGCTACGGTGTACAATGCTTCGCTAAATACCCAGAACATATGCGT CAGAATGACTTCTTCAAGTCAGCAATGCCTGATGGATATGTTCAAGAAAGAAC AATCTCTTTTAAAGAGGACGGAACTTATAAGACCAGAGCTGAAGTCAAATTCG AGGGAGAGGCCTTGGTGAACAGAATTGACTTAAAAGGACTGGAATTCAAGGA AGATGGCAATATCCTCGGTCACAAGCTTGAATACAGTTTTAATTCACACTACG TGTATATAACCGCGGATAAGAACAGGAATGGTTTAGAAGCTCAGTTTAGAATA AGACATAACGTGGACGATGGATCCGTGCAGCTAGCAGATCACTATCAGCAAA ATACGCCGATAGGGGAAGGCCCTGTTCTTTTGCCAGAGCAGCATTACCTAAC TACTAACTCTGTTTTGTCTAAGGACCCACAAGAGCGGAGAGATCATATGGTTT TGGTTGAGTTCGTTACGGCAGCTGGACTCTCCCTAGGCATGGATGAATTATA CAAATCAGGATCTGAGAATTTATATTTTCAAGGTCAATTGAGTTATGGATACGA CGAAAAATCAACTGGAATATCAGTGCCAGGACCAATGGGTCCATCAGGACCT AGGGGTCTTCCAGGGCCACCTGGCGCTCCTGGTCCTCAGGGATTTCAAGGT CCTCCAGGTGAACCTGGAGAGCCTGGTGCTTCTGGTCCAATGGGACCAAGA GGCCCTCCTGGTCCACCTGGTAAAAATGGTGATGATGGTGAGGCTGGAAAG CCTGGTCGTCCTGGAGAAAGAGGACCACCAGGACCACAGGGAGCTAGAGGT TTGCCAGGTACAGCCGGTCTTCCAGGAATGAAAGGACACAGGGGTTTTTCTG GTCTTGATGGTGCTAAGGGAGACGCAGGACCTGCTGGACCAAAGGGTGAAC CTGGATCTCCTGGTGAAAATGGTGCTCCTGGACAAATGGGACCTAGAGGATT GCCTGGCGAAAGAGGAAGACCTGGTGCTCCTGGACCAGCTGGAGCTAGAGG AAATGATGGAGCTACTGGTGCTGCCGGACCACCAGGACCAACTGGACCAGC TGGACCACCTGGTTTTCCAGGAGCAGTTGGGGCTAAGGGAGAAGGTGGACC ACAAGGACCTAGAGGATCAGAAGGACCTCAAGGAGTTAGGGGAGAACCAGG TCCTCCTGGTCCTGCTGGAGCTGCTGGACCCGCTGGTAACCCAGGAGCAGA TGGACAACCTGGAGCAAAAGGGGCTAATGGTGCTCCTGGTATTGCAGGTGCT CCTGGATTCCCTGGTGCTAGAGGACCTTCAGGACCTCAGGGACCATCCGGT CCTCCAGGACCAAAGGGAAACTCTGGTGAACCAGGGGCTCCTGGATCTAAG GGAGATACTGGAGCTAAGGGTGAGCCAGGACCTACAGGAATTCAAGGACCA CCAGGACCTGCAGGAGAGGAAGGTAAGAGGGGTGCTAGAGGTGAACCTGGA CCAGCTGGATTGCCAGGCCCACCTGGAGAGAGGGGTGGTCCAGGTTCTAGA GGTTTTCCAGGAGCTGATGGAGTTGCTGGTCCAAAGGGTCCTGCTGGTGAGA GAGGTGCACCTGGACCAGCAGGACCTAAAGGATCACCTGGAGAAGCTGGTC

[0791] GTCCTGGTGAGGCTGGTCTTCCAGGTGCTAAGGGCCTTACAGGATCTCCAG

[0792] GATCACCAGGTCCAGATGGAAAGACTGGACCACCAGGTCCTGCAGGACAAG

[0793] ATGGAAGGCCTGGTCCACCTGGACCACCTGGTGCTAGGGGACAAGCTGGTG

[0794] TTATGGGATTCCCAGGTCCAAAGGGTGCAGCTGGTGAGCCTGGTAAAGCTG

[0795] GTGAAAGAGGAGTTCCAGGACCTCCTGGTGCTGTTGGACCTGCTGGTAAAGA

[0796] TGGTGAAGCAGGTGCTCAGGGTCCACCAGGTCCAGCCGGTCCAGCTGGTGA

[0797] GAGGGGAGAGCAAGGTCCTGCTGGATCTCCTGGATTTCAAGGTTTGCCTGGA

[0798] CCTGCTGGACCTCCAGGAGAGGCTGGTAAACCTGGTGAGCAGGGTGTGCCT

[0799] GGTGATCTTGGTGCTCCTGGACCTAGTGGCGCAAGAGGCGAAAGGGGATTT

[0800] CCAGGTGAAAGAGGAGTTCAGGGACCACCAGGTCCAGCTGGTCCAAGAGGC

[0801] GCTAATGGAGCTCCAGGAAATGATGGAGCAAAGGGTGATGCTGGCGCTCCA

[0802] GGAGCTCCTGGTTCTCAAGGTGCACCTGGATTGCAAGGAATGCCAGGAGAG

[0803] AGAGGTGCAGCTGGTTTGCCGGGACCAAAGGGTGATAGAGGTGATGCTGGT

[0804] CCTAAGGGAGCTGATGGAGCTCCAGGAAAGGATGGTGTGAGAGGATTGACT

[0805] GGTCCAATTGGACCACCAGGTCCTGCTGGAGCACCAGGAGATAAAGGAGAA

[0806] GCTGGTCCTTCAGGACCAGCCGGTCCAACTGGAGCTAGAGGAGCTCCAGGA

[0807] GATAGAGGTGAACCAGGACCACCAGGACCTGCTGGGTTTGCAGGTCCACCT

[0808] GGTGCTGATGGACAACCTGGTGCTAAAGGAGAACCAGGCGATGCTGGTGCT

[0809] AAGGGAGATGCTGGCCCACCTGGTCCAGCTGGTCCTGCTGGGCCACCAGGT

[0810] CCTATCGGAAATGTTGGTGCTCCAGGGCCTAAAGGAGCTAGAGGATCAGCA

[0811] GGTCCCCCAGGTGCTACAGGTTTTCCAGGGGCTGCTGGAAGAGTGGGTCCC

[0812] CCAGGTCCTTCTGGGAATGCAGGACCACCAGGTCCTCCAGGACCAGCTGGA

[0813] AAAGAAGGAAGTAAGGGACCAAGGGGAGAAACTGGACCTGCAGGACGTCCA

[0814] GGTGAAGTTGGTCCACCTGGACCTCCAGGTCCAGCTGGAGAGAAAGGAGCT

[0815] CCAGGAGCTGATGGTCCAGCAGGTGCTCCTGGTACTCCAGGTCCACAAGGT

[0816] ATTGCTGGTCAAAGGGGAGTGGTTGGTCTCCCAGGTCAGAGAGGTGAAAGA

[0817] GGTTTCCCAGGTTTGCCTGGACCATCTGGAGAACCAGGTAAGCAGGGCCCAT

[0818] CTGGAGCTTCTGGTGAGAGAGGGCCACCTGGACCTATGGGACCTCCTGGGC

[0819] TTGCTGGACCTCCAGGGGAATCTGGTAGGGAGGGTGCTCCAGGTGCTGAAG

[0820] GATCACCTGGTAGAGATGGATCACCTGGAGCTAAAGGTGATAGAGGGGAAA

[0821] CAGGTCCAGCAGGTCCACCTGGCGCTCCTGGTGCTCCAGGAGCACCAGGGC

[0822] CAGTTGGACCTGCTGGAAAGTCTGGAGATAGGGGAGAGACTGGACCAGCTG

[0823] GACCTGCTGGACCAATCGGTCCTGTTGGCGCTAGAGGTCCTGCTGGACCAC

[0824] AAGGTCCTAGGGGAGATAAAGGTGAAACAGGTGAACAAGGAGATAGAGGAA

[0825] TTAAAGGTCATAGAGGGTTTTCTGGTTTGCAAGGACCTCCAGGACCTCCTGG

[0826] ATCTCCAGGAGAGCAAGGACCATCTGGTGCAAGCGGACCTGCTGGTCCTAG

[0827] AGGACCTCCAGGTTCAGCTGGCTCACCTGGTAAGGATGGATTAAATGGTTTG CCTGGACCAATAGGTCCACCAGGACCTAGAGGAAGGACAGGTGATGCAGGA

[0828] CCAGCTGGTCCACCTGGTCCACCAGGACCACCAGGACCACCAGGTCCACCT

[0829] TCTGGTGGATATGATCTTAGTTTTCTTCCTCAACCTCCCCAGGAGAAAGCTCA

[0830] TGATGGTGGTAGATATTATAGGGCTGCATTTGTTTATTAAACCCAGCTTTCTTG

[0831] TACAAAGTGGTAGCTCGAATTTCCCCGATCGTTCAAACATTTGGCAATAAAGT

[0832] TTCTTAAGATTGAATCCTGTTGCCGGTCTTGCGATGATTATCATATAATTTCTG

[0833] TTGAATTACGTTAAGCATGTAATAATTAACATGTAATGCATGACGTTATTTATG

[0834] AGATGGGTTTTTATGATTAGAGTCCCGCAATTATACATTTAATACGCGATAGAA

[0835] AACAAAATATAGCGCGCAAACTAGGATAAATTATCGCGCGCGGTGTCATCTAT

[0836] GTTACTAGATCGGGAAAGCTTTTAAACTATCAGTGTTTGACAGGATATATTGG

[0837] CGGGTAAACCTAAGAGAAAAGAGCGTTTATTAGAATAATCGGATATTTAAAAG

[0838] GGCGTGAAAAGGTTTATCCGTTCGTCCATTTGTATGTGCATGCCAACCACAG

[0839] GGTTCCCCTCGGGATCAAAGTACTTTGATCCAACCCCTCCGCTGCTATAGTG

[0840] CAGTCGGCTTCTGACGTTCAGTGCAGCCGTCTTCTGAAAACGACATGTCGCA

[0841] CAAGTCCTAAGTTACGCGACAGGCTGCCGCCCTGCCCTTTTCCTGGCGTTTT

[0842] CTTGTCGCGTGTTTTAGTCGCATAAAGTAGAATACTTGCGACTAGAACCGGAG

[0843] ACATTACGCCATGAACAAGAGCGCCGCCGCTGGCCTGCTGGGCTATGCCCG

[0844] CGTCAGCACCGACGACCAGGACTTGACCAACCAACGGGCCGAACTGCACGC

[0845] GGCCGGCTGCACCAAGCTGTTTTCCGAGAAGATCACCGGCACCAGGCGCGA

[0846] CCGCCCGGAGCTGGCCAGGATGCTTGACCACCTACGCCCTGGCGACGTTGT

[0847] GACAGTGACCAGGCTAGACCGCCTGGCCCGCAGCACCCGCGACCTACTGGA

[0848] CATTGCCGAGCGCATCCAGGAGGCCGGCGCGGGCCTGCGTAGCCTGGCAG

[0849] AGCCGTGGGCCGACACCACCACGCCGGCCGGCCGCATGGTGTTGACCGTG

[0850] TTCGCCGGCATTGCCGAGTTCGAGCGTTCCCTAATCATCGACCGCACCCGGA

[0851] GCGGGCGCGAGGCCGCCAAGGCCCGAGGCGTGAAGTTTGGCCCCCGCCCT

[0852] ACCCTCACCCCGGCACAGATCGCGCACGCCCGCGAGCTGATCGACCAGGAA

[0853] GGCCGCACCGTGAAAGAGGCGGCTGCACTGCTTGGCGTGCATCGCTCGACC

[0854] CTGTACCGCGCACTTGAGCGCAGCGAGGAAGTGACGCCCACCGAGGCCAG

[0855] GCGGCGCGGTGCCTTCCGTGAGGACGCATTGACCGAGGCCGACGCCCTGG

[0856] CGGCCGCCGAGAATGAACGCCAAGAGGAACAAGCATGAAACCGCACCAGGA

[0857] CGGCCAGGACGAACCGTTTTTCATTACCGAAGAGATCGAGGCGGAGATGATC

[0858] GCGGCCGGGTACGTGTTCGAGCCGCCCGCGCACGTCTCAACCGTGCGGCT

[0859] GCATGAAATCCTGGCCGGTTTGTCTGATGCCAAGCTGGCGGCCTGGCCGGC

[0860] CAGCTTGGCCGCTGAAGAAACCGAGCGCCGCCGTCTAAAAAGGTGATGTGT

[0861] ATTTGAGTAAAACAGCTTGCGTCATGCGGTCGCTGCGTATATGATGCGATGA

[0862] GTAAATAAACAAATACGCAAGGGGAACGCATGAAGGTTATCGCTGTACTTAAC

[0863] CAGAAAGGCGGGTCAGGCAAGACGACCATCGCAACCCATCTAGCCCGCGCC

[0864] CTGCAACTCGCCGGGGCCGATGTTCTGTTAGTCGATTCCGATCCCCAGGGCA GTGCCCGCGATTGGGCGGCCGTGCGGGAAGATCAACCGCTAACCGTTGTCG

[0865] GCATCGACCGCCCGACGATTGACCGCGACGTGAAGGCCATCGGCCGGCGC

[0866] GACTTCGTAGTGATCGACGGAGCGCCCCAGGCGGCGGACTTGGCTGTGTCC

[0867] GCGATCAAGGCAGCCGACTTCGTGCTGATTCCGGTGCAGCCAAGCCCTTAC

[0868] GACATATGGGCCACCGCCGACCTGGTGGAGCTGGTTAAGCAGCGCATTGAG

[0869] GTCACGGATGGAAGGCTACAAGCGGCCTTTGTCGTGTCGCGGGCGATCAAA

[0870] GGCACGCGCATCGGCGGTGAGGTTGCCGAGGCGCTGGCCGGGTACGAGCT

[0871] GCCCATTCTTGAGTCCCGTATCACGCAGCGCGTGAGCTACCCAGGCACTGC

[0872] CGCCGCCGGCACAACCGTTCTTGAATCAGAACCCGAGGGCGACGCTGCCCG

[0873] CGAGGTCCAGGCGCTGGCCGCTGAAATTAAATCAAAACTCATTTGAGTTAAT

[0874] GAGGTAAAGAGAAAATGAGCAAAAGCACAAACACGCTAAGTGCCGGCCGTC

[0875] CGAGCGCACGCAGCAGCAAGGCTGCAACGTTGGCCAGCCTGGCAGACACG

[0876] CCAGCCATGAAGCGGGTCAACTTTCAGTTGCCGGCGGAGGATCACACCAAG

[0877] CTGAAGATGTACGCGGTACGCCAAGGCAAGACCATTACCGAGCTGCTATCTG

[0878] AATACATCGCGCAGCTACCAGAGTAAATGAGCAAATGAATAAATGAGTAGATG

[0879] AATTTTAGCGGCTAAAGGAGGCGGCATGGAAAATCAAGAACAACCAGGCACC

[0880] GACGCCGTGGAATGCCCCATGTGTGGAGGAACGGGCGGTTGGCCAGGCGT

[0881] AAGCGGCTGGGTTGCCTGCCGGCCCTGCAATGGCACTGGAACCCCCAAGCC

[0882] CGAGGAATCGGCGTGAGCGGTCGCAAACCATCCGGCCCGGTACAAATCGGC

[0883] GCGGCGCTGGGTGATGACCTGGTGGAGAAGTTGAAGGCCGCGCAGGCCGC

[0884] CCAGCGGCAACGCATCGAGGCAGAAGCACGCCCCGGTGAATCGTGGCAAG

[0885] CGGCCGCTGATCGAATCCGCAAAGAATCCCGGCAACCGCCGGCAGCCGGTG

[0886] CGCCGTCGATTAGGAAGCCGCCCAAGGGCGACGAGCAACCAGATTTTTTCGT

[0887] TCCGATGCTCTATGACGTGGGCACCCGCGATAGTCGCAGCATCATGGACGT

[0888] GGCCGTTTTCCGTCTGTCGAAGCGTGACCGACGAGCTGGCGAGGTGATCCG

[0889] CTACGAGCTTCCAGACGGGCACGTAGAGGTTTCCGCAGGGCCGGCCGGCAT

[0890] GGCCAGTGTGTGGGATTACGACCTGGTACTGATGGCGGTTTCCCATCTAACC

[0891] GAATCCATGAACCGATACCGGGAAGGGAAGGGAGACAAGCCCGGCCGCGT

[0892] GTTCCGTCCACACGTTGCGGACGTACTCAAGTTCTGCCGGCGAGCCGATGG

[0893] CGGAAAGCAGAAAGACGACCTGGTAGAAACCTGCATTCGGTTAAACACCACG

[0894] CACGTTGCCATGCAGCGTACGAAGAAGGCCAAGAACGGCCGCCTGGTGACG

[0895] GTATCCGAGGGTGAAGCCTTGATTAGCCGCTACAAGATCGTAAAGAGCGAAA

[0896] CCGGGCGGCCGGAGTACATCGAGATCGAGCTAGCTGATTGGATGTACCGCG

[0897] AGATCACAGAAGGCAAGAACCCGGACGTGCTGACGGTTCACCCCGATTACTT

[0898] TTTGATCGATCCCGGCATCGGCCGTTTTCTCTACCGCCTGGCACGCCGCGCC

[0899] GCAGGCAAGGCAGAAGCCAGATGGTTGTTCAAGACGATCTACGAACGCAGT

[0900] GGCAGCGCCGGAGAGTTCAAGAAGTTCTGTTTCACCGTGCGCAAGCTGATC

[0901] GGGTCAAATGACCTGCCGGAGTACGATTTGAAGGAGGAGGCGGGGCAGGCT GGCCCGATCCTAGTCATGCGCTACCGCAACCTGATCGAGGGCGAAGCATCC GCCGGTTCCTAATGTACGGAGCAGATGCTAGGGCAAATTGCCCTAGCAGGG

[0902] GAAAAAGGTCGAAAAGGTCTCTTTCCTGTGGATAGCACGTACATTGGGAACC CAAAGCCGTACATTGGGAACCGGAACCCGTACATTGGGAACCCAAAGCCGTA CATTGGGAACCGGTCACACATGTAAGTGACTGATATAAAAGAGAAAAAAGGC GATTTTTCCGCCTAAAACTCTTTAAAACTTATTAAAACTCTTAAAACCCGCCTG GCCTGTGCATAACTGTCTGGCCAGCGCACAGCCGAAGAGCTGCAAAAAGCG CCTACCCTTCGGTCGCTGCGCTCCCTACGCCCCGCCGCTTCGCGTCGGCCT ATCGCGGCCGCTGGCCGCTCAAAAATGGCTGGCCTACGGCCAGGCAATCTA CCAGGGCGCGGACAAGCCGCGCCGTCGCCACTCGACCGCCGGCGCCCACA

[0903] TCAAGGCACCCTGCCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACA CATGCAGCTCCCGGAGACGGTCACAGCTTGTCTGTAAGCGGATGCCGGGAG CAGACAAGCCCGTCAGGGCGCGTCAGCGGGTGTTGGCGGGTGTCGGGGCG CAGCCATGACCCAGTCACGTAGCGATAGCGGAGTGTATACTGGCTTAACTAT GCGGCATCAGAGCAGATTGTACTGAGAGTGCACCATATGCGGTGTGAAATAC CGCACAGATGCGTAAGGAGAAAATACCGCATCAGGCGCTCTTCCGCTTCCTC GCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGC TCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGA

[0904] AAGAACATGTGAGCAAAAGGCCAGCAAA

[0905] Bovine Collagen alpha-1 (I) Expression vector 10;

[0906] SEQ ID NO: 21 :

[0907] AGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCG CCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAA CCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTG CGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCC CTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTC GGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCA GCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTA AGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAG

[0908] CGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGG CTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCT TCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAG CGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTC AAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAAC TCACGTTAAGGGATTTTGGTCATGCATTCTAGGTACTAAAACAATTCATCCAG TAAAATATAATATTTTATTTTCTCCCAATCAGGCTTGATCCCCAGTAAGTCAAA AAATAGCTCGACATACTGTTCTTCCCCGATATCCTCCCTGATCGACCGGACG CAGAAGGCAATGTCATACCACTTGTCCGCCCTGCCGCTTCTCCCAAGATCAA

[0909] TAAAGCCACTTACTTTGCCATCTTTCACAAAGATGTTGCTGTCTCCCAGGTCG

[0910] CCGTGGGAAAAGACAAGTTCCTCTTCGGGCTTTTCCGTCTTTAAAAAATCATA

[0911] CAGCTCGCGCGGATCTTTAAATGGAGTGTCTTCTTCCCAGTTTTCGCAATCCA

[0912] CATCGGCCAGATCGTTATTCAGTAAGTAATCCAATTCGGCTAAGCGGCTGTCT

[0913] AAGCTATTCGTATAGGGACAATCCGATATGTCGATGGAGTGAAAGAGCCTGA

[0914] TGCACTCCGCATACAGCTCGATAATCTTTTCAGGGCTTTGTTCATCTTCATACT

[0915] CTTCCGAGCAAAGGACGCCATCGGCCTCACTCATGAGCAGATTGCTCCAGCC

[0916] ATCATGCCGTTCAAAGTGCAGGACCTTTGGAACAGGCAGCTTTCCTTCCAGC

[0917] CATAGCATCATGTCCTTTTCCCGTTCCACATCATAGGTGGTCCCTTTATACCG

[0918] GCTGTCCGTCATTTTTAAATATAGGTTTTCATTTTCTCCCACCAGCTTATATAC

[0919] CTTAGCAGGAGACATTCCTTCCGTATCTTTTACGCAGCGGTATTTTTCGATCA

[0920] GTTTTTTCAATTCCGGTGATATTCTCATTTTAGCCATTTATTATTTCCTTCCTCT

[0921] TTTCTACAGTATTTAAAGATACCCCAAGAAGCTAATTATAACAAGACGAACTCC

[0922] AATTCACTGTTCCTTGCATTCTAAAACCTTAAATACCAGAAAACAGCTTTTTCA

[0923] AAGTTGTTTTCAAAGTTGGCGTATAACATAGTATCGACGGAGCCGATTTTGAA

[0924] ACCGCGGTGATCACAGGCAGCAACGCTCTGTCATCGTTACAATCAACATGCT

[0925] ACCCTCCGCGAGATCATCCGTGTTTCAAACCCGGCAGCTTAGTTGCCGTTCT

[0926] TCCGAATAGCATCGGTAACATGAGCAAAGTCTGCCGCCTTACAACGGCTCTC

[0927] CCGCTGACGCCGTCCCGGACTGATGGGCTGCCTGTATCGAGTGGTGATTTT

[0928] GTGCCGAGCTGCCGGTCGGGGAGCTGTTGGCTGGCTGGTGGCAGGATATAT

[0929] TGTGGTGTAAACAAATTGACGCTTAGACAACTTAATAACACATTGCGGACGTT

[0930] TTTAATGTACTGAATTAACGCCGAATTGAATTCGAGCTCGGTGGAGGATCTGG

[0931] ATTTTAGTACTGGATTTTGGTTTTAGGAATTAGAAATTTTATTGATAGAAGTATT

[0932] TTACAAATACAAATACATACTAAGGGTTTCTTATATGCTCAACACATGAGCGAA

[0933] ACCCTATAGGAACCCTAATTCCCTTATCTGGGAACTACTCACACATTATTATG

[0934] GAGAAACTATTCACTCTCCTTCTCAAACGTTTCTGTTCCCTCTGGACAGCCCC

[0935] GACTGACGTTGGACTCAACCTCAATGGGCGCTAATTGTAATAGCTCCTGTTTG

[0936] CTTCTTATGGCCATTTCGCAAAGGTGTTGCAGTGTTCTGCTACCACCACTCAC

[0937] TGTAATTGACACACCTCTGAAACGGATGGAGTAAGTGCAACCAATTTGATCAA

[0938] ATCCAAAGAAACGACTAGCAGCGTAATTTACTGAGTCACCAGTCCAGGAACC

[0939] TAGAACTCGATGGAGAGAGGCCTCTGTCCGATCATAGCGTAGGTAGCGTTTG

[0940] AAAACTACCTTGCCAAATCCCCAGGACTCTTTAAATCCCAGTGGGTTGTCCTG

[0941] GTTGGAGTTTGTTTCGTCATTGTGTAAACGCCATTCTGTCCAAGATGGGCTCT

[0942] CATCGGGTAACTTAAATGGGCTGGTAGTGCCGCCAGAACCACCGTCCCATCT

[0943] CTCTGAATTGGCCTGCTCTCTTGCATCATTACCCTGGATTGCTCTCTCCATTG

[0944] TCCTCTCCAAATGAAATGAACTTCCTTATATAGAGGAAGGGTCTTGCGAAGGA

[0945] TAGTGGGATTGTGCGTCATCCCTTACGTCAGTGGAGATGTCACATCAATCCA CTTGCTTTGTAGACGTGGTTGGAACCTCTTCTTTTTCCACGATGCTCCTCGTG GGTGGGGGTCCATCTTTGGGACCACTGTCGGCAGAGAGATCTTGAATGATAG CCTTTCCTTTATCGCAATGATGGCATTTGTAGGAGCCACCTTCCTTTTCTACT GTCCTTTCGATGAAGTGACAGATAGCTGGGCAATGGAATCCGAGGAGGTTTC CCGAAATTATCCTTTGTTGAAAAGTCTCAAACAATAGGCAACCGTGGACTTCT TCACTAGCTCATCGAGATAGCAATGCCACTAGCTAATTTCTTACGTTGTATCTT ATTTGTTTTACTTTGGGGCATGACATGGTTAAACCCCATCAAAGAGAAAGTGT TTCATCCACTAGTCAATATACGAATTCATTCGAATTTATCCCTGTAAATCCTAG TTCTTAGGATGAACTGGTGTAATAAACAGCAAAAAAAAATAAAAAAAAATAAAT CTGGAATCATTCGACCACCTCAATAAACTAAAGCTACCAATTACCACAATATA GTC ATC C ATATC C ACTTAGATATAAAAGATAAAAGTAAAC AAATATTAAATTTC ATATGCACGCATAGGAAACTCATGATCTTATCTTTTAAATAGACATCTAGTTTT CTTAGGTTATAAATAGACATTTTGTCCTAGAACTTCTTCACTACTAAAACCTAG CCTCAAAAAGCAGGCTGCCACCATGGTGAGCAGTGGTGAGGACATCTTTTCA GGCCTAGTCCCGATACTTATTGAACTTGAAGGAGACGTCAATGGGCACAGAT TCTCAGTACGAGGCGAGGGATACGGGGATGCTTCCAATGGAAAGCTTGAGAT AAAATTTATATGTACAACTGGAAGGCTTCCTGTCCCATGGCCAACTTTAGTTA CCACTCTTAGCTACGGTGTACAATGCTTCGCTAAATACCCAGAACATATGCGT CAGAATGACTTCTTCAAGTCAGCAATGCCTGATGGATATGTTCAAGAAAGAAC AATCTCTTTTAAAGAGGACGGAACTTATAAGACCAGAGCTGAAGTCAAATTCG AGGGAGAGGCCTTGGTGAACAGAATTGACTTAAAAGGACTGGAATTCAAGGA AGATGGCAATATCCTCGGTCACAAGCTTGAATACAGTTTTAATTCACACTACG TGTATATAACCGCGGATAAGAACAGGAATGGTTTAGAAGCTCAGTTTAGAATA AGACATAACGTGGACGATGGATCCGTGCAGCTAGCAGATCACTATCAGCAAA ATACGCCGATAGGGGAAGGCCCTGTTCTTTTGCCAGAGCAGCATTACCTAAC TACTAACTCTGTTTTGTCTAAGGACCCACAAGAGCGGAGAGATCATATGGTTT TGGTTGAGTTCGTTACGGCAGCTGGACTCTCCCTAGGCATGGATGAATTATA CAAATCAGGATCTGAGAATTTATATTTTCAAGGTCAATTGAGTTATGGATACGA CGAAAAATCAACTGGAATATCAGTGCCAGGACCAATGGGTCCATCAGGACCT AGGGGTCTTCCAGGGCCACCTGGCGCTCCTGGTCCTCAGGGATTTCAAGGT CCTCCAGGTGAACCTGGAGAGCCTGGTGCTTCTGGTCCAATGGGACCAAGA

[0946] GGCCCTCCTGGTCCACCTGGTAAAAATGGTGATGATGGTGAGGCTGGAAAG CCTGGTCGTCCTGGAGAAAGAGGACCACCAGGACCACAGGGAGCTAGAGGT TTGCCAGGTACAGCCGGTCTTCCAGGAATGAAAGGACACAGGGGTTTTTCTG GTCTTGATGGTGCTAAGGGAGACGCAGGACCTGCTGGACCAAAGGGTGAAC CTGGATCTCCTGGTGAAAATGGTGCTCCTGGACAAATGGGACCTAGAGGATT GCCTGGCGAAAGAGGAAGACCTGGTGCTCCTGGACCAGCTGGAGCTAGAGG AAATGATGGAGCTACTGGTGCTGCCGGACCACCAGGACCAACTGGACCAGC TGGACCACCTGGTTTTCCAGGAGCAGTTGGGGCTAAGGGAGAAGGTGGACC

[0947] ACAAGGACCTAGAGGATCAGAAGGACCTCAAGGAGTTAGGGGAGAACCAGG

[0948] TCCTCCTGGTCCTGCTGGAGCTGCTGGACCCGCTGGTAACCCAGGAGCAGA

[0949] TGGACAACCTGGAGCAAAAGGGGCTAATGGTGCTCCTGGTATTGCAGGTGCT

[0950] CCTGGATTCCCTGGTGCTAGAGGACCTTCAGGACCTCAGGGACCATCCGGT

[0951] CCTCCAGGACCAAAGGGAAACTCTGGTGAACCAGGGGCTCCTGGATCTAAG

[0952] GGAGATACTGGAGCTAAGGGTGAGCCAGGACCTACAGGAATTCAAGGACCA

[0953] CCAGGACCTGCAGGAGAGGAAGGTAAGAGGGGTGCTAGAGGTGAACCTGGA

[0954] CCAGCTGGATTGCCAGGCCCACCTGGAGAGAGGGGTGGTCCAGGTTCTAGA

[0955] GGTTTTCCAGGAGCTGATGGAGTTGCTGGTCCAAAGGGTCCTGCTGGTGAGA

[0956] GAGGTGCACCTGGACCAGCAGGACCTAAAGGATCACCTGGAGAAGCTGGTC

[0957] GTCCTGGTGAGGCTGGTCTTCCAGGTGCTAAGGGCCTTACAGGATCTCCAG

[0958] GATCACCAGGTCCAGATGGAAAGACTGGACCACCAGGTCCTGCAGGACAAG

[0959] ATGGAAGGCCTGGTCCACCTGGACCACCTGGTGCTAGGGGACAAGCTGGTG

[0960] TTATGGGATTCCCAGGTCCAAAGGGTGCAGCTGGTGAGCCTGGTAAAGCTG

[0961] GTGAAAGAGGAGTTCCAGGACCTCCTGGTGCTGTTGGACCTGCTGGTAAAGA

[0962] TGGTGAAGCAGGTGCTCAGGGTCCACCAGGTCCAGCCGGTCCAGCTGGTGA

[0963] GAGGGGAGAGCAAGGTCCTGCTGGATCTCCTGGATTTCAAGGTTTGCCTGGA

[0964] CCTGCTGGACCTCCAGGAGAGGCTGGTAAACCTGGTGAGCAGGGTGTGCCT

[0965] GGTGATCTTGGTGCTCCTGGACCTAGTGGCGCAAGAGGCGAAAGGGGATTT

[0966] CCAGGTGAAAGAGGAGTTCAGGGACCACCAGGTCCAGCTGGTCCAAGAGGC

[0967] GCTAATGGAGCTCCAGGAAATGATGGAGCAAAGGGTGATGCTGGCGCTCCA

[0968] GGAGCTCCTGGTTCTCAAGGTGCACCTGGATTGCAAGGAATGCCAGGAGAG

[0969] AGAGGTGCAGCTGGTTTGCCGGGACCAAAGGGTGATAGAGGTGATGCTGGT

[0970] CCTAAGGGAGCTGATGGAGCTCCAGGAAAGGATGGTGTGAGAGGATTGACT

[0971] GGTCCAATTGGACCACCAGGTCCTGCTGGAGCACCAGGAGATAAAGGAGAA

[0972] GCTGGTCCTTCAGGACCAGCCGGTCCAACTGGAGCTAGAGGAGCTCCAGGA

[0973] GATAGAGGTGAACCAGGACCACCAGGACCTGCTGGGTTTGCAGGTCCACCT

[0974] GGTGCTGATGGACAACCTGGTGCTAAAGGAGAACCAGGCGATGCTGGTGCT

[0975] AAGGGAGATGCTGGCCCACCTGGTCCAGCTGGTCCTGCTGGGCCACCAGGT

[0976] CCTATCGGAAATGTTGGTGCTCCAGGGCCTAAAGGAGCTAGAGGATCAGCA

[0977] GGTCCCCCAGGTGCTACAGGTTTTCCAGGGGCTGCTGGAAGAGTGGGTCCC

[0978] CCAGGTCCTTCTGGGAATGCAGGACCACCAGGTCCTCCAGGACCAGCTGGA

[0979] AAAGAAGGAAGTAAGGGACCAAGGGGAGAAACTGGACCTGCAGGACGTCCA

[0980] GGTGAAGTTGGTCCACCTGGACCTCCAGGTCCAGCTGGAGAGAAAGGAGCT

[0981] CCAGGAGCTGATGGTCCAGCAGGTGCTCCTGGTACTCCAGGTCCACAAGGT

[0982] ATTGCTGGTCAAAGGGGAGTGGTTGGTCTCCCAGGTCAGAGAGGTGAAAGA

[0983] GGTTTCCCAGGTTTGCCTGGACCATCTGGAGAACCAGGTAAGCAGGGCCCAT CTGGAGCTTCTGGTGAGAGAGGGCCACCTGGACCTATGGGACCTCCTGGGC

[0984] TTGCTGGACCTCCAGGGGAATCTGGTAGGGAGGGTGCTCCAGGTGCTGAAG

[0985] GATCACCTGGTAGAGATGGATCACCTGGAGCTAAAGGTGATAGAGGGGAAA

[0986] CAGGTCCAGCAGGTCCACCTGGCGCTCCTGGTGCTCCAGGAGCACCAGGGC

[0987] CAGTTGGACCTGCTGGAAAGTCTGGAGATAGGGGAGAGACTGGACCAGCTG

[0988] GACCTGCTGGACCAATCGGTCCTGTTGGCGCTAGAGGTCCTGCTGGACCAC

[0989] AAGGTCCTAGGGGAGATAAAGGTGAAACAGGTGAACAAGGAGATAGAGGAA

[0990] TTAAAGGTCATAGAGGGTTTTCTGGTTTGCAAGGACCTCCAGGACCTCCTGG

[0991] ATCTCCAGGAGAGCAAGGACCATCTGGTGCAAGCGGACCTGCTGGTCCTAG

[0992] AGGACCTCCAGGTTCAGCTGGCTCACCTGGTAAGGATGGATTAAATGGTTTG

[0993] CCTGGACCAATAGGTCCACCAGGACCTAGAGGAAGGACAGGTGATGCAGGA

[0994] CCAGCTGGTCCACCTGGTCCACCAGGACCACCAGGACCACCAGGTCCACCT

[0995] TCTGGTGGATATGATCTTAGTTTTCTTCCTCAACCTCCCCAGGAGAAAGCTCA

[0996] TGATGGTGGTAGATATTATAGGGCTTAAACCCAGCTTTCTTGTACAAAGTGGT

[0997] AGCTCGAATTTCCCCGATCGTTCAAACATTTGGCAATAAAGTTTCTTAAGATTG

[0998] AATCCTGTTGCCGGTCTTGCGATGATTATCATATAATTTCTGTTGAATTACGTT

[0999] AAGCATGTAATAATTAACATGTAATGCATGACGTTATTTATGAGATGGGTTTTT

[1000] ATGATTAGAGTCCCGCAATTATACATTTAATACGCGATAGAAAACAAAATATAG

[1001] CGCGCAAACTAGGATAAATTATCGCGCGCGGTGTCATCTATGTTACTAGATC

[1002] GGGAAAGCTTTTAAACTATCAGTGTTTGACAGGATATATTGGCGGGTAAACCT

[1003] AAGAGAAAAGAGCGTTTATTAGAATAATCGGATATTTAAAAGGGCGTGAAAAG

[1004] GTTTATCCGTTCGTCCATTTGTATGTGCATGCCAACCACAGGGTTCCCCTCGG

[1005] GATCAAAGTACTTTGATCCAACCCCTCCGCTGCTATAGTGCAGTCGGCTTCTG

[1006] ACGTTCAGTGCAGCCGTCTTCTGAAAACGACATGTCGCACAAGTCCTAAGTTA

[1007] CGCGACAGGCTGCCGCCCTGCCCTTTTCCTGGCGTTTTCTTGTCGCGTGTTT

[1008] TAGTCGCATAAAGTAGAATACTTGCGACTAGAACCGGAGACATTACGCCATG

[1009] AACAAGAGCGCCGCCGCTGGCCTGCTGGGCTATGCCCGCGTCAGCACCGAC

[1010] GACCAGGACTTGACCAACCAACGGGCCGAACTGCACGCGGCCGGCTGCACC

[1011] AAGCTGTTTTCCGAGAAGATCACCGGCACCAGGCGCGACCGCCCGGAGCTG

[1012] GCCAGGATGCTTGACCACCTACGCCCTGGCGACGTTGTGACAGTGACCAGG

[1013] CTAGACCGCCTGGCCCGCAGCACCCGCGACCTACTGGACATTGCCGAGCGC

[1014] ATCCAGGAGGCCGGCGCGGGCCTGCGTAGCCTGGCAGAGCCGTGGGCCGA

[1015] CACCACCACGCCGGCCGGCCGCATGGTGTTGACCGTGTTCGCCGGCATTGC

[1016] CGAGTTCGAGCGTTCCCTAATCATCGACCGCACCCGGAGCGGGCGCGAGGC

[1017] CGCCAAGGCCCGAGGCGTGAAGTTTGGCCCCCGCCCTACCCTCACCCCGGC

[1018] ACAGATCGCGCACGCCCGCGAGCTGATCGACCAGGAAGGCCGCACCGTGAA

[1019] AGAGGCGGCTGCACTGCTTGGCGTGCATCGCTCGACCCTGTACCGCGCACT

[1020] TGAGCGCAGCGAGGAAGTGACGCCCACCGAGGCCAGGCGGCGCGGTGCCT TCCGTGAGGACGCATTGACCGAGGCCGACGCCCTGGCGGCCGCCGAGAAT

[1021] GAACGCCAAGAGGAACAAGCATGAAACCGCACCAGGACGGCCAGGACGAAC

[1022] CGTTTTTCATTACCGAAGAGATCGAGGCGGAGATGATCGCGGCCGGGTACGT

[1023] GTTCGAGCCGCCCGCGCACGTCTCAACCGTGCGGCTGCATGAAATCCTGGC

[1024] CGGTTTGTCTGATGCCAAGCTGGCGGCCTGGCCGGCCAGCTTGGCCGCTGA

[1025] AGAAACCGAGCGCCGCCGTCTAAAAAGGTGATGTGTATTTGAGTAAAACAGC

[1026] TTGCGTCATGCGGTCGCTGCGTATATGATGCGATGAGTAAATAAACAAATAC

[1027] GCAAGGGGAACGCATGAAGGTTATCGCTGTACTTAACCAGAAAGGCGGGTC

[1028] AGGCAAGACGACCATCGCAACCCATCTAGCCCGCGCCCTGCAACTCGCCGG

[1029] GGCCGATGTTCTGTTAGTCGATTCCGATCCCCAGGGCAGTGCCCGCGATTG

[1030] GGCGGCCGTGCGGGAAGATCAACCGCTAACCGTTGTCGGCATCGACCGCCC

[1031] GACGATTGACCGCGACGTGAAGGCCATCGGCCGGCGCGACTTCGTAGTGAT

[1032] CGACGGAGCGCCCCAGGCGGCGGACTTGGCTGTGTCCGCGATCAAGGCAG

[1033] CCGACTTCGTGCTGATTCCGGTGCAGCCAAGCCCTTACGACATATGGGCCAC

[1034] CGCCGACCTGGTGGAGCTGGTTAAGCAGCGCATTGAGGTCACGGATGGAAG

[1035] GCTACAAGCGGCCTTTGTCGTGTCGCGGGCGATCAAAGGCACGCGCATCGG

[1036] CGGTGAGGTTGCCGAGGCGCTGGCCGGGTACGAGCTGCCCATTCTTGAGTC

[1037] CCGTATCACGCAGCGCGTGAGCTACCCAGGCACTGCCGCCGCCGGCACAAC

[1038] CGTTCTTGAATCAGAACCCGAGGGCGACGCTGCCCGCGAGGTCCAGGCGCT

[1039] GGCCGCTGAAATTAAATCAAAACTCATTTGAGTTAATGAGGTAAAGAGAAAAT

[1040] GAGCAAAAGCACAAACACGCTAAGTGCCGGCCGTCCGAGCGCACGCAGCAG

[1041] CAAGGCTGCAACGTTGGCCAGCCTGGCAGACACGCCAGCCATGAAGCGGGT

[1042] CAACTTTCAGTTGCCGGCGGAGGATCACACCAAGCTGAAGATGTACGCGGTA

[1043] CGCCAAGGCAAGACCATTACCGAGCTGCTATCTGAATACATCGCGCAGCTAC

[1044] CAGAGTAAATGAGCAAATGAATAAATGAGTAGATGAATTTTAGCGGCTAAAGG

[1045] AGGCGGCATGGAAAATCAAGAACAACCAGGCACCGACGCCGTGGAATGCCC

[1046] CATGTGTGGAGGAACGGGCGGTTGGCCAGGCGTAAGCGGCTGGGTTGCCT

[1047] GCCGGCCCTGCAATGGCACTGGAACCCCCAAGCCCGAGGAATCGGCGTGA

[1048] GCGGTCGCAAACCATCCGGCCCGGTACAAATCGGCGCGGCGCTGGGTGATG

[1049] ACCTGGTGGAGAAGTTGAAGGCCGCGCAGGCCGCCCAGCGGCAACGCATC

[1050] GAGGCAGAAGCACGCCCCGGTGAATCGTGGCAAGCGGCCGCTGATCGAATC

[1051] CGCAAAGAATCCCGGCAACCGCCGGCAGCCGGTGCGCCGTCGATTAGGAAG

[1052] CCGCCCAAGGGCGACGAGCAACCAGATTTTTTCGTTCCGATGCTCTATGACG

[1053] TGGGCACCCGCGATAGTCGCAGCATCATGGACGTGGCCGTTTTCCGTCTGTC

[1054] GAAGCGTGACCGACGAGCTGGCGAGGTGATCCGCTACGAGCTTCCAGACGG

[1055] GCACGTAGAGGTTTCCGCAGGGCCGGCCGGCATGGCCAGTGTGTGGGATTA

[1056] CGACCTGGTACTGATGGCGGTTTCCCATCTAACCGAATCCATGAACCGATAC

[1057] CGGGAAGGGAAGGGAGACAAGCCCGGCCGCGTGTTCCGTCCACACGTTGC GGACGTACTCAAGTTCTGCCGGCGAGCCGATGGCGGAAAGCAGAAAGACGA

[1058] CCTGGTAGAAACCTGCATTCGGTTAAACACCACGCACGTTGCCATGCAGCGT

[1059] ACGAAGAAGGCCAAGAACGGCCGCCTGGTGACGGTATCCGAGGGTGAAGCC

[1060] TTGATTAGCCGCTACAAGATCGTAAAGAGCGAAACCGGGCGGCCGGAGTAC

[1061] ATCGAGATCGAGCTAGCTGATTGGATGTACCGCGAGATCACAGAAGGCAAGA

[1062] ACCCGGACGTGCTGACGGTTCACCCCGATTACTTTTTGATCGATCCCGGCAT

[1063] CGGCCGTTTTCTCTACCGCCTGGCACGCCGCGCCGCAGGCAAGGCAGAAGC

[1064] CAGATGGTTGTTCAAGACGATCTACGAACGCAGTGGCAGCGCCGGAGAGTT

[1065] CAAGAAGTTCTGTTTCACCGTGCGCAAGCTGATCGGGTCAAATGACCTGCCG

[1066] GAGTACGATTTGAAGGAGGAGGCGGGGCAGGCTGGCCCGATCCTAGTCATG

[1067] CGCTACCGCAACCTGATCGAGGGCGAAGCATCCGCCGGTTCCTAATGTACG

[1068] GAGCAGATGCTAGGGCAAATTGCCCTAGCAGGGGAAAAAGGTCGAAAAGGT

[1069] CTCTTTCCTGTGGATAGCACGTACATTGGGAACCCAAAGCCGTACATTGGGA

[1070] ACCGGAACCCGTACATTGGGAACCCAAAGCCGTACATTGGGAACCGGTCAC

[1071] ACATGTAAGTGACTGATATAAAAGAGAAAAAAGGCGATTTTTCCGCCTAAAAC

[1072] TCTTTAAAACTTATTAAAACTCTTAAAACCCGCCTGGCCTGTGCATAACTGTCT

[1073] GGCCAGCGCACAGCCGAAGAGCTGCAAAAAGCGCCTACCCTTCGGTCGCTG

[1074] CGCTCCCTACGCCCCGCCGCTTCGCGTCGGCCTATCGCGGCCGCTGGCCG

[1075] CTCAAAAATGGCTGGCCTACGGCCAGGCAATCTACCAGGGCGCGGACAAGC

[1076] CGCGCCGTCGCCACTCGACCGCCGGCGCCCACATCAAGGCACCCTGCCTCG

[1077] CGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCGGAGAC

[1078] GGTCACAGCTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGCCCGTCAGGG

[1079] CGCGTCAGCGGGTGTTGGCGGGTGTCGGGGCGCAGCCATGACCCAGTCAC

[1080] GTAGCGATAGCGGAGTGTATACTGGCTTAACTATGCGGCATCAGAGCAGATT

[1081] GTACTGAGAGTGCACCATATGCGGTGTGAAATACCGCACAGATGCGTAAGGA

[1082] GAAAATACCGCATCAGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCG

[1083] CTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAAT

[1084] ACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAA

[1085] GGCCAGCAAA

[1086] Bovine Collagen alpha-1 (I) Expression vector 11 ;

[1087] SEQ ID NO: 25:

[1088] AGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCG

[1089] CCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAA

[1090] CCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTG

[1091] CGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCC

[1092] CTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTC

[1093] GGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCA GCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTA

[1094] AGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAG

[1095] CGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGG

[1096] CTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCT

[1097] TCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAG

[1098] CGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTC

[1099] AAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAAC

[1100] TCACGTTAAGGGATTTTGGTCATGCATTCTAGGTACTAAAACAATTCATCCAG

[1101] TAAAATATAATATTTTATTTTCTCCCAATCAGGCTTGATCCCCAGTAAGTCAAA

[1102] AAATAGCTCGACATACTGTTCTTCCCCGATATCCTCCCTGATCGACCGGACG

[1103] CAGAAGGCAATGTCATACCACTTGTCCGCCCTGCCGCTTCTCCCAAGATCAA

[1104] TAAAGCCACTTACTTTGCCATCTTTCACAAAGATGTTGCTGTCTCCCAGGTCG

[1105] CCGTGGGAAAAGACAAGTTCCTCTTCGGGCTTTTCCGTCTTTAAAAAATCATA

[1106] CAGCTCGCGCGGATCTTTAAATGGAGTGTCTTCTTCCCAGTTTTCGCAATCCA

[1107] CATCGGCCAGATCGTTATTCAGTAAGTAATCCAATTCGGCTAAGCGGCTGTCT

[1108] AAGCTATTCGTATAGGGACAATCCGATATGTCGATGGAGTGAAAGAGCCTGA

[1109] TGCACTCCGCATACAGCTCGATAATCTTTTCAGGGCTTTGTTCATCTTCATACT

[1110] CTTCCGAGCAAAGGACGCCATCGGCCTCACTCATGAGCAGATTGCTCCAGCC

[1111] ATCATGCCGTTCAAAGTGCAGGACCTTTGGAACAGGCAGCTTTCCTTCCAGC

[1112] CATAGCATCATGTCCTTTTCCCGTTCCACATCATAGGTGGTCCCTTTATACCG

[1113] GCTGTCCGTCATTTTTAAATATAGGTTTTCATTTTCTCCCACCAGCTTATATAC

[1114] CTTAGCAGGAGACATTCCTTCCGTATCTTTTACGCAGCGGTATTTTTCGATCA

[1115] GTTTTTTCAATTCCGGTGATATTCTCATTTTAGCCATTTATTATTTCCTTCCTCT

[1116] TTTCTACAGTATTTAAAGATACCCCAAGAAGCTAATTATAACAAGACGAACTCC

[1117] AATTCACTGTTCCTTGCATTCTAAAACCTTAAATACCAGAAAACAGCTTTTTCA

[1118] AAGTTGTTTTCAAAGTTGGCGTATAACATAGTATCGACGGAGCCGATTTTGAA

[1119] ACCGCGGTGATCACAGGCAGCAACGCTCTGTCATCGTTACAATCAACATGCT

[1120] ACCCTCCGCGAGATCATCCGTGTTTCAAACCCGGCAGCTTAGTTGCCGTTCT

[1121] TCCGAATAGCATCGGTAACATGAGCAAAGTCTGCCGCCTTACAACGGCTCTC

[1122] CCGCTGACGCCGTCCCGGACTGATGGGCTGCCTGTATCGAGTGGTGATTTT

[1123] GTGCCGAGCTGCCGGTCGGGGAGCTGTTGGCTGGCTGGTGGCAGGATATAT

[1124] TGTGGTGTAAACAAATTGACGCTTAGACAACTTAATAACACATTGCGGACGTT

[1125] TTTAATGTACTGAATTAACGCCGAATTGAATTCGAGCTCGGTGGAGGATCTGG

[1126] ATTTTAGTACTGGATTTTGGTTTTAGGAATTAGAAATTTTATTGATAGAAGTATT

[1127] TTACAAATACAAATACATACTAAGGGTTTCTTATATGCTCAACACATGAGCGAA

[1128] ACCCTATAGGAACCCTAATTCCCTTATCTGGGAACTACTCACACATTATTATG

[1129] GAGAAACTATTCACTCTCCTTCTCAAACGTTTCTGTTCCCTCTGGACAGCCCC

[1130] GACTGACGTTGGACTCAACCTCAATGGGCGCTAATTGTAATAGCTCCTGTTTG CTTCTTATGGCCATTTCGCAAAGGTGTTGCAGTGTTCTGCTACCACCACTCAC TGTAATTGACACACCTCTGAAACGGATGGAGTAAGTGCAACCAATTTGATCAA ATCCAAAGAAACGACTAGCAGCGTAATTTACTGAGTCACCAGTCCAGGAACC TAGAACTCGATGGAGAGAGGCCTCTGTCCGATCATAGCGTAGGTAGCGTTTG AAAACTACCTTGCCAAATCCCCAGGACTCTTTAAATCCCAGTGGGTTGTCCTG GTTGGAGTTTGTTTCGTCATTGTGTAAACGCCATTCTGTCCAAGATGGGCTCT CATCGGGTAACTTAAATGGGCTGGTAGTGCCGCCAGAACCACCGTCCCATCT CTCTGAATTGGCCTGCTCTCTTGCATCATTACCCTGGATTGCTCTCTCCATTG TCCTCTCCAAATGAAATGAACTTCCTTATATAGAGGAAGGGTCTTGCGAAGGA TAGTGGGATTGTGCGTCATCCCTTACGTCAGTGGAGATGTCACATCAATCCA CTTGCTTTGTAGACGTGGTTGGAACCTCTTCTTTTTCCACGATGCTCCTCGTG GGTGGGGGTCCATCTTTGGGACCACTGTCGGCAGAGAGATCTTGAATGATAG CCTTTCCTTTATCGCAATGATGGCATTTGTAGGAGCCACCTTCCTTTTCTACT GTCCTTTCGATGAAGTGACAGATAGCTGGGCAATGGAATCCGAGGAGGTTTC CCGAAATTATCCTTTGTTGAAAAGTCTCAAACAATAGGCAACCGTGGACTTCT TCACTAGCTCATCGAGATAGCAATGCCACTAGCTAATTTCTTACGTTGTATCTT ATTTGTTTTACTTTGGGGCATGACATGGTTAAACCCCATCAAAGAGAAAGTGT TTCATCCACTAGTCAATATACGAATTCATTCGAATTTATCCCTGTAAATCCTAG TTCTTAGGATGAACTGGTGTAATAAACAGCAAAAAAAAATAAAAAAAAATAAAT CTGGAATCATTCGACCACCTCAATAAACTAAAGCTACCAATTACCACAATATA

[1131] GTC ATC C ATATC C ACTTAGATATAAAAGATAAAAGTAAAC AAATATTAAATTTC ATATGCACGCATAGGAAACTCATGATCTTATCTTTTAAATAGACATCTAGTTTT CTTAGGTTATAAATAGACATTTTGTCCTAGAACTTCTTCACTACTAAAACCTAG CCTCAAAAAGCAGGCTGCCACCATGGTGAGCAGTGGTGAGGACATCTTTTCA GGCCTAGTCCCGATACTTATTGAACTTGAAGGAGACGTCAATGGGCACAGAT TCTCAGTACGAGGCGAGGGATACGGGGATGCTTCCAATGGAAAGCTTGAGAT AAAATTTATATGTACAACTGGAAGGCTTCCTGTCCCATGGCCAACTTTAGTTA CCACTCTTAGCTACGGTGTACAATGCTTCGCTAAATACCCAGAACATATGCGT CAGAATGACTTCTTCAAGTCAGCAATGCCTGATGGATATGTTCAAGAAAGAAC AATCTCTTTTAAAGAGGACGGAACTTATAAGACCAGAGCTGAAGTCAAATTCG AGGGAGAGGCCTTGGTGAACAGAATTGACTTAAAAGGACTGGAATTCAAGGA AGATGGCAATATCCTCGGTCACAAGCTTGAATACAGTTTTAATTCACACTACG TGTATATAACCGCGGATAAGAACAGGAATGGTTTAGAAGCTCAGTTTAGAATA AGACATAACGTGGACGATGGATCCGTGCAGCTAGCAGATCACTATCAGCAAA ATACGCCGATAGGGGAAGGCCCTGTTCTTTTGCCAGAGCAGCATTACCTAAC TACTAACTCTGTTTTGTCTAAGGACCCACAAGAGCGGAGAGATCATATGGTTT TGGTTGAGTTCGTTACGGCAGCTGGACTCTCCCTAGGCATGGATGAATTATA CAAATCAGGATCTGAGAATTTATATTTTCAAGGTCAATTGAGTTATGGATACGA CGAAAAATCAACTGGAATATCAGTGCCAGGACCAATGGGTCCATCAGGACCT

[1132] AGGGGTCTTCCAGGGCCACCTGGCGCTCCTGGTCCTCAGGGATTTCAAGGT

[1133] CCTCCAGGTGAACCTGGAGAGCCTGGTGCTTCTGGTCCAATGGGACCAAGA

[1134] GGCCCTCCTGGTCCACCTGGTAAAAATGGTGATGATGGTGAGGCTGGAAAG

[1135] CCTGGTCGTCCTGGAGAAAGAGGACCACCAGGACCACAGGGAGCTAGAGGT

[1136] TTGCCAGGTACAGCCGGTCTTCCAGGAATGAAAGGACACAGGGGTTTTTCTG

[1137] GTCTTGATGGTGCTAAGGGAGACGCAGGACCTGCTGGACCAAAGGGTGAAC

[1138] CTGGATCTCCTGGTGAAAATGGTGCTCCTGGACAAATGGGACCTAGAGGATT

[1139] GCCTGGCGAAAGAGGAAGACCTGGTGCTCCTGGACCAGCTGGAGCTAGAGG

[1140] AAATGATGGAGCTACTGGTGCTGCCGGACCACCAGGACCAACTGGACCAGC

[1141] TGGACCACCTGGTTTTCCAGGAGCAGTTGGGGCTAAGGGAGAAGGTGGACC

[1142] ACAAGGACCTAGAGGATCAGAAGGACCTCAAGGAGTTAGGGGAGAACCAGG

[1143] TCCTCCTGGTCCTGCTGGAGCTGCTGGACCCGCTGGTAACCCAGGAGCAGA

[1144] TGGACAACCTGGAGCAAAAGGGGCTAATGGTGCTCCTGGTATTGCAGGTGCT

[1145] CCTGGATTCCCTGGTGCTAGAGGACCTTCAGGACCTCAGGGACCATCCGGT

[1146] CCTCCAGGACCAAAGGGAAACTCTGGTGAACCAGGGGCTCCTGGATCTAAG

[1147] GGAGATACTGGAGCTAAGGGTGAGCCAGGACCTACAGGAATTCAAGGACCA

[1148] CCAGGACCTGCAGGAGAGGAAGGTAAGAGGGGTGCTAGAGGTGAACCTGGA

[1149] CCAGCTGGATTGCCAGGCCCACCTGGAGAGAGGGGTGGTCCAGGTTCTAGA

[1150] GGTTTTCCAGGAGCTGATGGAGTTGCTGGTCCAAAGGGTCCTGCTGGTGAGA

[1151] GAGGTGCACCTGGACCAGCAGGACCTAAAGGATCACCTGGAGAAGCTGGTC

[1152] GTCCTGGTGAGGCTGGTCTTCCAGGTGCTAAGGGCCTTACAGGATCTCCAG

[1153] GATCACCAGGTCCAGATGGAAAGACTGGACCACCAGGTCCTGCAGGACAAG

[1154] ATGGAAGGCCTGGTCCACCTGGACCACCTGGTGCTAGGGGACAAGCTGGTG

[1155] TTATGGGATTCCCAGGTCCAAAGGGTGCAGCTGGTGAGCCTGGTAAAGCTG

[1156] GTGAAAGAGGAGTTCCAGGACCTCCTGGTGCTGTTGGACCTGCTGGTAAAGA

[1157] TGGTGAAGCAGGTGCTCAGGGTCCACCAGGTCCAGCCGGTCCAGCTGGTGA

[1158] GAGGGGAGAGCAAGGTCCTGCTGGATCTCCTGGATTTCAAGGTTTGCCTGGA

[1159] CCTGCTGGACCTCCAGGAGAGGCTGGTAAACCTGGTGAGCAGGGTGTGCCT

[1160] GGTGATCTTGGTGCTCCTGGACCTAGTGGCGCAAGAGGCGAAAGGGGATTT

[1161] CCAGGTGAAAGAGGAGTTCAGGGACCACCAGGTCCAGCTGGTCCAAGAGGC

[1162] GCTAATGGAGCTCCAGGAAATGATGGAGCAAAGGGTGATGCTGGCGCTCCA

[1163] GGAGCTCCTGGTTCTCAAGGTGCACCTGGATTGCAAGGAATGCCAGGAGAG

[1164] AGAGGTGCAGCTGGTTTGCCGGGACCAAAGGGTGATAGAGGTGATGCTGGT

[1165] CCTAAGGGAGCTGATGGAGCTCCAGGAAAGGATGGTGTGAGAGGATTGACT

[1166] GGTCCAATTGGACCACCAGGTCCTGCTGGAGCACCAGGAGATAAAGGAGAA

[1167] GCTGGTCCTTCAGGACCAGCCGGTCCAACTGGAGCTAGAGGAGCTCCAGGA

[1168] GATAGAGGTGAACCAGGACCACCAGGACCTGCTGGGTTTGCAGGTCCACCT GGTGCTGATGGACAACCTGGTGCTAAAGGAGAACCAGGCGATGCTGGTGCT

[1169] AAGGGAGATGCTGGCCCACCTGGTCCAGCTGGTCCTGCTGGGCCACCAGGT

[1170] CCTATCGGAAATGTTGGTGCTCCAGGGCCTAAAGGAGCTAGAGGATCAGCA

[1171] GGTCCCCCAGGTGCTACAGGTTTTCCAGGGGCTGCTGGAAGAGTGGGTCCC

[1172] CCAGGTCCTTCTGGGAATGCAGGACCACCAGGTCCTCCAGGACCAGCTGGA

[1173] AAAGAAGGAAGTAAGGGACCAAGGGGAGAAACTGGACCTGCAGGACGTCCA

[1174] GGTGAAGTTGGTCCACCTGGACCTCCAGGTCCAGCTGGAGAGAAAGGAGCT

[1175] CCAGGAGCTGATGGTCCAGCAGGTGCTCCTGGTACTCCAGGTCCACAAGGT

[1176] ATTGCTGGTCAAAGGGGAGTGGTTGGTCTCCCAGGTCAGAGAGGTGAAAGA

[1177] GGTTTCCCAGGTTTGCCTGGACCATCTGGAGAACCAGGTAAGCAGGGCCCAT

[1178] CTGGAGCTTCTGGTGAGAGAGGGCCACCTGGACCTATGGGACCTCCTGGGC

[1179] TTGCTGGACCTCCAGGGGAATCTGGTAGGGAGGGTGCTCCAGGTGCTGAAG

[1180] GATCACCTGGTAGAGATGGATCACCTGGAGCTAAAGGTGATAGAGGGGAAA

[1181] CAGGTCCAGCAGGTCCACCTGGCGCTCCTGGTGCTCCAGGAGCACCAGGGC

[1182] CAGTTGGACCTGCTGGAAAGTCTGGAGATAGGGGAGAGACTGGACCAGCTG

[1183] GACCTGCTGGACCAATCGGTCCTGTTGGCGCTAGAGGTCCTGCTGGACCAC

[1184] AAGGTCCTAGGGGAGATAAAGGTGAAACAGGTGAACAAGGAGATAGAGGAA

[1185] TTAAAGGTCATAGAGGGTTTTCTGGTTTGCAAGGACCTCCAGGACCTCCTGG

[1186] ATCTCCAGGAGAGCAAGGACCATCTGGTGCAAGCGGACCTGCTGGTCCTAG

[1187] AGGACCTCCAGGTTCAGCTGGCTCACCTGGTAAGGATGGATTAAATGGTTTG

[1188] CCTGGACCAATAGGTCCACCAGGACCTAGAGGAAGGACAGGTGATGCAGGA

[1189] CCAGCTGGTCCACCTGGTCCACCAGGACCACCAGGACCACCAGGTCCACCT

[1190] TCTGGTGGATATGATCTTAGTTTTCTTCCTCAACCTCCCCAGGAGAAAGCTCA

[1191] TGATGGTGGTAGATATTATAGGGCTAAGGATGAGCTTTAAACCCAGCTTTCTT

[1192] GTACAAAGTGGTAGCTCGAATTTCCCCGATCGTTCAAACATTTGGCAATAAAG

[1193] >

[1194] TTTCTTAAGATTGAATCCTGTTGCCGGTCTTGCGATGATTATCATATAATTTCT

[1195] GTTGAATTACGTTAAGCATGTAATAATTAACATGTAATGCATGACGTTATTTAT

[1196] GAGATGGGTTTTTATGATTAGAGTCCCGCAATTATACATTTAATACGCGATAG

[1197] AAAACAAAATATAGCGCGCAAACTAGGATAAATTATCGCGCGCGGTGTCATCT

[1198] ATGTTACTAGATCGGGAAAGCTTTTAAACTATCAGTGTTTGACAGGATATATTG

[1199] GCGGGTAAACCTAAGAGAAAAGAGCGTTTATTAGAATAATCGGATATTTAAAA

[1200] GGGCGTGAAAAGGTTTATCCGTTCGTCCATTTGTATGTGCATGCCAACCACA

[1201] GGGTTCCCCTCGGGATCAAAGTACTTTGATCCAACCCCTCCGCTGCTATAGT

[1202] GCAGTCGGCTTCTGACGTTCAGTGCAGCCGTCTTCTGAAAACGACATGTCGC

[1203] ACAAGTCCTAAGTTACGCGACAGGCTGCCGCCCTGCCCTTTTCCTGGCGTTT

[1204] TCTTGTCGCGTGTTTTAGTCGCATAAAGTAGAATACTTGCGACTAGAACCGGA

[1205] GACATTACGCCATGAACAAGAGCGCCGCCGCTGGCCTGCTGGGCTATGCCC

[1206] GCGTCAGCACCGACGACCAGGACTTGACCAACCAACGGGCCGAACTGCACG CGGCCGGCTGCACCAAGCTGTTTTCCGAGAAGATCACCGGCACCAGGCGCG

[1207] ACCGCCCGGAGCTGGCCAGGATGCTTGACCACCTACGCCCTGGCGACGTTG

[1208] TGACAGTGACCAGGCTAGACCGCCTGGCCCGCAGCACCCGCGACCTACTGG

[1209] ACATTGCCGAGCGCATCCAGGAGGCCGGCGCGGGCCTGCGTAGCCTGGCA

[1210] GAGCCGTGGGCCGACACCACCACGCCGGCCGGCCGCATGGTGTTGACCGT

[1211] GTTCGCCGGCATTGCCGAGTTCGAGCGTTCCCTAATCATCGACCGCACCCG

[1212] GAGCGGGCGCGAGGCCGCCAAGGCCCGAGGCGTGAAGTTTGGCCCCCGCC

[1213] CTACCCTCACCCCGGCACAGATCGCGCACGCCCGCGAGCTGATCGACCAGG

[1214] AAGGCCGCACCGTGAAAGAGGCGGCTGCACTGCTTGGCGTGCATCGCTCGA

[1215] CCCTGTACCGCGCACTTGAGCGCAGCGAGGAAGTGACGCCCACCGAGGCCA

[1216] GGCGGCGCGGTGCCTTCCGTGAGGACGCATTGACCGAGGCCGACGCCCTG

[1217] GCGGCCGCCGAGAATGAACGCCAAGAGGAACAAGCATGAAACCGCACCAGG

[1218] ACGGCCAGGACGAACCGTTTTTCATTACCGAAGAGATCGAGGCGGAGATGAT

[1219] CGCGGCCGGGTACGTGTTCGAGCCGCCCGCGCACGTCTCAACCGTGCGGC

[1220] TGCATGAAATCCTGGCCGGTTTGTCTGATGCCAAGCTGGCGGCCTGGCCGG

[1221] CCAGCTTGGCCGCTGAAGAAACCGAGCGCCGCCGTCTAAAAAGGTGATGTG

[1222] TATTTGAGTAAAACAGCTTGCGTCATGCGGTCGCTGCGTATATGATGCGATGA

[1223] GTAAATAAACAAATACGCAAGGGGAACGCATGAAGGTTATCGCTGTACTTAAC

[1224] CAGAAAGGCGGGTCAGGCAAGACGACCATCGCAACCCATCTAGCCCGCGCC

[1225] CTGCAACTCGCCGGGGCCGATGTTCTGTTAGTCGATTCCGATCCCCAGGGCA

[1226] GTGCCCGCGATTGGGCGGCCGTGCGGGAAGATCAACCGCTAACCGTTGTCG

[1227] GCATCGACCGCCCGACGATTGACCGCGACGTGAAGGCCATCGGCCGGCGC

[1228] GACTTCGTAGTGATCGACGGAGCGCCCCAGGCGGCGGACTTGGCTGTGTCC

[1229] GCGATCAAGGCAGCCGACTTCGTGCTGATTCCGGTGCAGCCAAGCCCTTAC

[1230] GACATATGGGCCACCGCCGACCTGGTGGAGCTGGTTAAGCAGCGCATTGAG

[1231] GTCACGGATGGAAGGCTACAAGCGGCCTTTGTCGTGTCGCGGGCGATCAAA

[1232] GGCACGCGCATCGGCGGTGAGGTTGCCGAGGCGCTGGCCGGGTACGAGCT

[1233] GCCCATTCTTGAGTCCCGTATCACGCAGCGCGTGAGCTACCCAGGCACTGC

[1234] CGCCGCCGGCACAACCGTTCTTGAATCAGAACCCGAGGGCGACGCTGCCCG

[1235] CGAGGTCCAGGCGCTGGCCGCTGAAATTAAATCAAAACTCATTTGAGTTAAT

[1236] GAGGTAAAGAGAAAATGAGCAAAAGCACAAACACGCTAAGTGCCGGCCGTC

[1237] CGAGCGCACGCAGCAGCAAGGCTGCAACGTTGGCCAGCCTGGCAGACACG

[1238] CCAGCCATGAAGCGGGTCAACTTTCAGTTGCCGGCGGAGGATCACACCAAG

[1239] CTGAAGATGTACGCGGTACGCCAAGGCAAGACCATTACCGAGCTGCTATCTG

[1240] AATACATCGCGCAGCTACCAGAGTAAATGAGCAAATGAATAAATGAGTAGATG

[1241] AATTTTAGCGGCTAAAGGAGGCGGCATGGAAAATCAAGAACAACCAGGCACC

[1242] GACGCCGTGGAATGCCCCATGTGTGGAGGAACGGGCGGTTGGCCAGGCGT

[1243] AAGCGGCTGGGTTGCCTGCCGGCCCTGCAATGGCACTGGAACCCCCAAGCC CGAGGAATCGGCGTGAGCGGTCGCAAACCATCCGGCCCGGTACAAATCGGC

[1244] GCGGCGCTGGGTGATGACCTGGTGGAGAAGTTGAAGGCCGCGCAGGCCGC

[1245] CCAGCGGCAACGCATCGAGGCAGAAGCACGCCCCGGTGAATCGTGGCAAG

[1246] CGGCCGCTGATCGAATCCGCAAAGAATCCCGGCAACCGCCGGCAGCCGGTG

[1247] CGCCGTCGATTAGGAAGCCGCCCAAGGGCGACGAGCAACCAGATTTTTTCGT

[1248] TCCGATGCTCTATGACGTGGGCACCCGCGATAGTCGCAGCATCATGGACGT

[1249] GGCCGTTTTCCGTCTGTCGAAGCGTGACCGACGAGCTGGCGAGGTGATCCG

[1250] CTACGAGCTTCCAGACGGGCACGTAGAGGTTTCCGCAGGGCCGGCCGGCAT

[1251] GGCCAGTGTGTGGGATTACGACCTGGTACTGATGGCGGTTTCCCATCTAACC

[1252] GAATCCATGAACCGATACCGGGAAGGGAAGGGAGACAAGCCCGGCCGCGT

[1253] GTTCCGTCCACACGTTGCGGACGTACTCAAGTTCTGCCGGCGAGCCGATGG

[1254] CGGAAAGCAGAAAGACGACCTGGTAGAAACCTGCATTCGGTTAAACACCACG

[1255] CACGTTGCCATGCAGCGTACGAAGAAGGCCAAGAACGGCCGCCTGGTGACG

[1256] GTATCCGAGGGTGAAGCCTTGATTAGCCGCTACAAGATCGTAAAGAGCGAAA

[1257] CCGGGCGGCCGGAGTACATCGAGATCGAGCTAGCTGATTGGATGTACCGCG

[1258] AGATCACAGAAGGCAAGAACCCGGACGTGCTGACGGTTCACCCCGATTACTT

[1259] TTTGATCGATCCCGGCATCGGCCGTTTTCTCTACCGCCTGGCACGCCGCGCC

[1260] GCAGGCAAGGCAGAAGCCAGATGGTTGTTCAAGACGATCTACGAACGCAGT

[1261] GGCAGCGCCGGAGAGTTCAAGAAGTTCTGTTTCACCGTGCGCAAGCTGATC

[1262] GGGTCAAATGACCTGCCGGAGTACGATTTGAAGGAGGAGGCGGGGCAGGCT

[1263] GGCCCGATCCTAGTCATGCGCTACCGCAACCTGATCGAGGGCGAAGCATCC

[1264] GCCGGTTCCTAATGTACGGAGCAGATGCTAGGGCAAATTGCCCTAGCAGGG

[1265] GAAAAAGGTCGAAAAGGTCTCTTTCCTGTGGATAGCACGTACATTGGGAACC

[1266] CAAAGCCGTACATTGGGAACCGGAACCCGTACATTGGGAACCCAAAGCCGTA

[1267] CATTGGGAACCGGTCACACATGTAAGTGACTGATATAAAAGAGAAAAAAGGC

[1268] GATTTTTCCGCCTAAAACTCTTTAAAACTTATTAAAACTCTTAAAACCCGCCTG

[1269] GCCTGTGCATAACTGTCTGGCCAGCGCACAGCCGAAGAGCTGCAAAAAGCG

[1270] CCTACCCTTCGGTCGCTGCGCTCCCTACGCCCCGCCGCTTCGCGTCGGCCT

[1271] ATCGCGGCCGCTGGCCGCTCAAAAATGGCTGGCCTACGGCCAGGCAATCTA

[1272] CCAGGGCGCGGACAAGCCGCGCCGTCGCCACTCGACCGCCGGCGCCCACA

[1273] TCAAGGCACCCTGCCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACA

[1274] CATGCAGCTCCCGGAGACGGTCACAGCTTGTCTGTAAGCGGATGCCGGGAG

[1275] CAGACAAGCCCGTCAGGGCGCGTCAGCGGGTGTTGGCGGGTGTCGGGGCG

[1276] CAGCCATGACCCAGTCACGTAGCGATAGCGGAGTGTATACTGGCTTAACTAT

[1277] GCGGCATCAGAGCAGATTGTACTGAGAGTGCACCATATGCGGTGTGAAATAC

[1278] CGCACAGATGCGTAAGGAGAAAATACCGCATCAGGCGCTCTTCCGCTTCCTC

[1279] GCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGC TCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGA

[1280] AAGAACATGTGAGCAAAAGGCCAGCAAA

[1281] Prolyl 4-hydroxylase expression vector P4HA_1 ;

[1282] SEQ ID NO: 29:

[1283] CAACTTTGTATAGAAAAGTTGGCCAACATGGTGGAGCACGACACTCTCGTCT

[1284] ACTCCAAGAATATCAAAGATACAGTCTCAGAAGACCAAAGGGCTATTGAGACT

[1285] TTTCAACAAAGGGTAATATCGGGAAACCTCCTCGGATTCCATTGCCCAGCTAT

[1286] CTGTCACTTCATCAAAAGGACAGTAGAAAAGGAAGGTGGCACCTACAAATGC

[1287] CATCATTGCGATAAAGGAAAGGCTATCGTTCAAGATGCCTCTGCCGACAGTG

[1288] GTCCCAAAGATGGACCCCCACCCACGAGGAGCATCGTGGAAAAAGAAGACG

[1289] TTCCAACCACGTCTTCAAAGCAAGTGGATTGATGTGAACATGGTGGAGCACG

[1290] ACACTCTCGTCTACTCCAAGAATATCAAAGATACAGTCTCAGAAGACCAAAGG

[1291] GCTATTGAGACTTTTCAACAAAGGGTAATATCGGGAAACCTCCTCGGATTCCA

[1292] TTGCCCAGCTATCTGTCACTTCATCAAAAGGACAGTAGAAAAGGAAGGTGGC

[1293] ACCTACAAATGCCATCATTGCGATAAAGGAAAGGCTATCGTTCAAGATGCCTC

[1294] TGCCGACAGTGGTCCCAAAGATGGACCCCCACCCACGAGGAGCATCGTGGA

[1295] AAAAGAAGACGTTCCAACCACGTCTTCAAAGCAAGTGGATTGATGTGATATCT

[1296] CCACTGACGTAAGGGATGACGCACAATCCCACTATCCTTCGCAAGACCCTTC

[1297] CTCTATATAAGGAAGTTCATTTCATTTGGAGAGGACACGCTGACAAGTTTGTA

[1298] CAAAAAAGCAGGCTGCCACCATGATTTGGTATATTTTGGTGGTTGGAATTTTA

[1299] TTGCCTCAATCTCTTGCACACCCCGGATTTTTTACCTCTATTGGACAAATGAC

[1300] GGATCTTATTCATACTGAAAAGGATTTGGTTACTAGCCTTAAAGATTACATTAA

[1301] AGCTGAGGAGGATAAACTTGAACAGATTAAGAAGTGGGCAGAAAAGTTGGAT

[1302] AGACTTACCTCTACTGCTACTAAGGATCCAGAGGGCTTCGTCGGACACCCTG

[1303] TTAACGCATTCAAGTTAATGAAGAGATTGAACACTGAATGGTCTGAACTTGAA

[1304] AATCTTGTGTTGAAGGACATGTCAGATGGTTTCATTTCTAATCTTACAATACAG

[1305] CGACAATATTTTCCAAACGATGAAGATCAAGTTGGCGCAGCTAAGGCACTTTT

[1306] GAGACTCCAAGATACTTATAACCTCGATACTGATACAATTTCTAAAGGTGATTT

[1307] GCCAGGTGTTAAGCATAAGAGTTTTTTGACTGTTGAAGATTGTTTCGAGCTTG

[1308] GTAAGGTTGCATATACTGAGGCTGATTACTATCATACTGAGTTGTGGATGGAA

[1309] CAAGCACTGAGGCAGCTAGATGAGGGTGAAGTTTCTACAGTTGACAAGGTTT

[1310] CTGTTCTTGATTACCTTTCTTATGCTGTTTATCAACAAGGAGATTTGGATAAAG

[1311] CTTTGCTTCTTACAAAAAAGCTTTTGGAACTTGATCCAGAACACCAGAGAGCT

[1312] AACGGTAATCTCAAATACTTTGAATATATTATGGCAAAAGAAAAAGATGCTAAC

[1313] AAGTCTTCATCTGATGATCAATCTGATCAAAAGACTACTCTTAAGAAGAAGGG

[1314] TGCTGCAGTTGATTATCTCCCTGAGAGACAGAAGTATGAAATGCTTTGTAGGG

[1315] GAGAAGGAATTAAAATGACACCTAGAAGACAGAAGAAACTTTTTTGTAGGTAT CATGATGGTAACAGGAATCCAAAATTTATTCTCGCTCCTGCTAAACAAGAGGA

[1316] TGAGTGGGACAAGCCAAGGATCATTCGTTTTCACGACATTATATCTGACGCTG

[1317] AGATCGAAGTTGTGAAGGATTTAGCTAAGCCTAGGCTTTCAAGAGCTACAGTT

[1318] CATGATCCAGAAACTGGAAAATTGACTACTGCTCAATACAGAGTTTCAAAGTC

[1319] TGCTTGGCTTTCAGGATATGAGAACCCTGTTGTGTCTAGGATTAATATGAGAA

[1320] TCCAAGATTTGACTGGACTTGATGTTTCAACTGCAGAAGAATTGCAGGTTGCT

[1321] AACTATGGTGTGGGAGGACAATATGAGCCTCATTTCGATTTTGCTCGGAAGG

[1322] ATGAACCAGATGCTTTTAAAGAACTCGGGACTGGTAACAGAATTGCCACTTG

[1323] GCTTTTTTATATGTCAGATGTGCTTGCTGGTGGTGCTACTGTTTTTCCTGAAGT

[1324] GGGAGCTTCTGTTTGGCCAAAGAAAGGTACCGCCGTCTTTTGGTATAATCTTT

[1325] TTGCAAGTGGTGAAGGTGACTATTCAACAAGGCATGCTGCATGCCCTGTGTT

[1326] GGTTGGTAATAAGTGGGTGTCTAATAAATGGCTTCATGAGCGTGGCCAGGAA

[1327] TTTAGAAGACCATGCACTCTGTCAGAGCTTGAAGCTTTTGTTTATTAGACCCA

[1328] GCTTTCTTGTACAAAGTGGTAGCTCGAATTTCCCCGATCGTTCAAACATTTGG

[1329] CAATAAAGTTTCTTAAGATTGAATCCTGTTGCCGGTCTTGCGATGATTATCATA

[1330] TAATTTCTGTTGAATTACGTTAAGCATGTAATAATTAACATGTAATGCATGACG

[1331] TTATTTATGAGATGGGTTTTTATGATTAGAGTCCCGCAATTATACATTTAATAC

[1332] GCGATAGAAAACAAAATATAGCGCGCAAACTAGGATAAATTATCGCGCGCGG

[1333] TGTCATCTATGTTACTAGATCGGGAAAGCTTTTAAACTATCAGTGTTTGACAG

[1334] GATATATTGGCGGGTAAACCTAAGAGAAAAGAGCGTTTATTAGAATAATCGGA

[1335] TATTTAAAAGGGCGTGAAAAGGTTTATCCGTTCGTCCATTTGTATGTGCATGC

[1336] CAACCACAGGGTTCCCCTCGGGATCAAAGTACTTTGATCCAACCCCTCCGCT

[1337] GCTATAGTGCAGTCGGCTTCTGACGTTCAGTGCAGCCGTCTTCTGAAAACGA

[1338] CATGTCGCACAAGTCCTAAGTTACGCGACAGGCTGCCGCCCTGCCCTTTTCC

[1339] TGGCGTTTTCTTGTCGCGTGTTTTAGTCGCATAAAGTAGAATACTTGCGACTA

[1340] GAACCGGAGACATTACGCCATGAACAAGAGCGCCGCCGCTGGCCTGCTGGG

[1341] CTATGCCCGCGTCAGCACCGACGACCAGGACTTGACCAACCAACGGGCCGA

[1342] ACTGCACGCGGCCGGCTGCACCAAGCTGTTTTCCGAGAAGATCACCGGCAC

[1343] CAGGCGCGACCGCCCGGAGCTGGCCAGGATGCTTGACCACCTACGCCCTG

[1344] GCGACGTTGTGACAGTGACCAGGCTAGACCGCCTGGCCCGCAGCACCCGCG

[1345] ACCTACTGGACATTGCCGAGCGCATCCAGGAGGCCGGCGCGGGCCTGCGTA

[1346] GCCTGGCAGAGCCGTGGGCCGACACCACCACGCCGGCCGGCCGCATGGTG

[1347] TTGACCGTGTTCGCCGGCATTGCCGAGTTCGAGCGTTCCCTAATCATCGACC

[1348] GCACCCGGAGCGGGCGCGAGGCCGCCAAGGCCCGAGGCGTGAAGTTTGGC

[1349] CCCCGCCCTACCCTCACCCCGGCACAGATCGCGCACGCCCGCGAGCTGATC

[1350] GACCAGGAAGGCCGCACCGTGAAAGAGGCGGCTGCACTGCTTGGCGTGCAT

[1351] CGCTCGACCCTGTACCGCGCACTTGAGCGCAGCGAGGAAGTGACGCCCACC

[1352] GAGGCCAGGCGGCGCGGTGCCTTCCGTGAGGACGCATTGACCGAGGCCGA CGCCCTGGCGGCCGCCGAGAATGAACGCCAAGAGGAACAAGCATGAAACCG

[1353] CACCAGGACGGCCAGGACGAACCGTTTTTCATTACCGAAGAGATCGAGGCG

[1354] GAGATGATCGCGGCCGGGTACGTGTTCGAGCCGCCCGCGCACGTCTCAACC

[1355] GTGCGGCTGCATGAAATCCTGGCCGGTTTGTCTGATGCCAAGCTGGCGGCC

[1356] TGGCCGGCCAGCTTGGCCGCTGAAGAAACCGAGCGCCGCCGTCTAAAAAGG

[1357] TGATGTGTATTTGAGTAAAACAGCTTGCGTCATGCGGTCGCTGCGTATATGAT

[1358] GCGATGAGTAAATAAACAAATACGCAAGGGGAACGCATGAAGGTTATCGCTG

[1359] TACTTAACCAGAAAGGCGGGTCAGGCAAGACGACCATCGCAACCCATCTAGC

[1360] CCGCGCCCTGCAACTCGCCGGGGCCGATGTTCTGTTAGTCGATTCCGATCC

[1361] CCAGGGCAGTGCCCGCGATTGGGCGGCCGTGCGGGAAGATCAACCGCTAA

[1362] CCGTTGTCGGCATCGACCGCCCGACGATTGACCGCGACGTGAAGGCCATCG

[1363] GCCGGCGCGACTTCGTAGTGATCGACGGAGCGCCCCAGGCGGCGGACTTG

[1364] GCTGTGTCCGCGATCAAGGCAGCCGACTTCGTGCTGATTCCGGTGCAGCCA

[1365] AGCCCTTACGACATATGGGCCACCGCCGACCTGGTGGAGCTGGTTAAGCAG

[1366] CGCATTGAGGTCACGGATGGAAGGCTACAAGCGGCCTTTGTCGTGTCGCGG

[1367] GCGATCAAAGGCACGCGCATCGGCGGTGAGGTTGCCGAGGCGCTGGCCGG

[1368] GTACGAGCTGCCCATTCTTGAGTCCCGTATCACGCAGCGCGTGAGCTACCCA

[1369] GGCACTGCCGCCGCCGGCACAACCGTTCTTGAATCAGAACCCGAGGGCGAC

[1370] GCTGCCCGCGAGGTCCAGGCGCTGGCCGCTGAAATTAAATCAAAACTCATTT

[1371] GAGTTAATGAGGTAAAGAGAAAATGAGCAAAAGCACAAACACGCTAAGTGCC

[1372] GGCCGTCCGAGCGCACGCAGCAGCAAGGCTGCAACGTTGGCCAGCCTGGC

[1373] AGACACGCCAGCCATGAAGCGGGTCAACTTTCAGTTGCCGGCGGAGGATCA

[1374] CACCAAGCTGAAGATGTACGCGGTACGCCAAGGCAAGACCATTACCGAGCT

[1375] GCTATCTGAATACATCGCGCAGCTACCAGAGTAAATGAGCAAATGAATAAATG

[1376] AGTAGATGAATTTTAGCGGCTAAAGGAGGCGGCATGGAAAATCAAGAACAAC

[1377] CAGGCACCGACGCCGTGGAATGCCCCATGTGTGGAGGAACGGGCGGTTGG

[1378] CCAGGCGTAAGCGGCTGGGTTGCCTGCCGGCCCTGCAATGGCACTGGAACC

[1379] CCCAAGCCCGAGGAATCGGCGTGAGCGGTCGCAAACCATCCGGCCCGGTAC

[1380] AAATCGGCGCGGCGCTGGGTGATGACCTGGTGGAGAAGTTGAAGGCCGCG

[1381] CAGGCCGCCCAGCGGCAACGCATCGAGGCAGAAGCACGCCCCGGTGAATC

[1382] GTGGCAAGCGGCCGCTGATCGAATCCGCAAAGAATCCCGGCAACCGCCGGC

[1383] AGCCGGTGCGCCGTCGATTAGGAAGCCGCCCAAGGGCGACGAGCAACCAG

[1384] ATTTTTTCGTTCCGATGCTCTATGACGTGGGCACCCGCGATAGTCGCAGCAT

[1385] CATGGACGTGGCCGTTTTCCGTCTGTCGAAGCGTGACCGACGAGCTGGCGA

[1386] GGTGATCCGCTACGAGCTTCCAGACGGGCACGTAGAGGTTTCCGCAGGGCC

[1387] GGCCGGCATGGCCAGTGTGTGGGATTACGACCTGGTACTGATGGCGGTTTC

[1388] CCATCTAACCGAATCCATGAACCGATACCGGGAAGGGAAGGGAGACAAGCC

[1389] CGGCCGCGTGTTCCGTCCACACGTTGCGGACGTACTCAAGTTCTGCCGGCG AGCCGATGGCGGAAAGCAGAAAGACGACCTGGTAGAAACCTGCATTCGGTT

[1390] AAACACCACGCACGTTGCCATGCAGCGTACGAAGAAGGCCAAGAACGGCCG

[1391] CCTGGTGACGGTATCCGAGGGTGAAGCCTTGATTAGCCGCTACAAGATCGTA

[1392] AAGAGCGAAACCGGGCGGCCGGAGTACATCGAGATCGAGCTAGCTGATTGG

[1393] ATGTACCGCGAGATCACAGAAGGCAAGAACCCGGACGTGCTGACGGTTCAC

[1394] CCCGATTACTTTTTGATCGATCCCGGCATCGGCCGTTTTCTCTACCGCCTGG

[1395] CACGCCGCGCCGCAGGCAAGGCAGAAGCCAGATGGTTGTTCAAGACGATCT

[1396] ACGAACGCAGTGGCAGCGCCGGAGAGTTCAAGAAGTTCTGTTTCACCGTGC

[1397] GCAAGCTGATCGGGTCAAATGACCTGCCGGAGTACGATTTGAAGGAGGAGG

[1398] CGGGGCAGGCTGGCCCGATCCTAGTCATGCGCTACCGCAACCTGATCGAGG

[1399] GCGAAGCATCCGCCGGTTCCTAATGTACGGAGCAGATGCTAGGGCAAATTG

[1400] CCCTAGCAGGGGAAAAAGGTCGAAAAGGTCTCTTTCCTGTGGATAGCACGTA

[1401] CATTGGGAACCCAAAGCCGTACATTGGGAACCGGAACCCGTACATTGGGAAC

[1402] CCAAAGCCGTACATTGGGAACCGGTCACACATGTAAGTGACTGATATAAAAG

[1403] AGAAAAAAGGCGATTTTTCCGCCTAAAACTCTTTAAAACTTATTAAAACTCTTA

[1404] AAACCCGCCTGGCCTGTGCATAACTGTCTGGCCAGCGCACAGCCGAAGAGC

[1405] TGCAAAAAGCGCCTACCCTTCGGTCGCTGCGCTCCCTACGCCCCGCCGCTT

[1406] CGCGTCGGCCTATCGCGGCCGCTGGCCGCTCAAAAATGGCTGGCCTACGGC

[1407] CAGGCAATCTACCAGGGCGCGGACAAGCCGCGCCGTCGCCACTCGACCGC

[1408] CGGCGCCCACATCAAGGCACCCTGCCTCGCGCGTTTCGGTGATGACGGTGA

[1409] AAACCTCTGACACATGCAGCTCCCGGAGACGGTCACAGCTTGTCTGTAAGCG

[1410] GATGCCGGGAGCAGACAAGCCCGTCAGGGCGCGTCAGCGGGTGTTGGCGG

[1411] GTGTCGGGGCGCAGCCATGACCCAGTCACGTAGCGATAGCGGAGTGTATAC

[1412] TGGCTTAACTATGCGGCATCAGAGCAGATTGTACTGAGAGTGCACCATATGC

[1413] GGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGCATCAGGCGCTC

[1414] TTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCG

[1415] AGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGG

[1416] GATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAAC

[1417] CGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACG

[1418] AGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACT

[1419] ATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTT

[1420] CCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCG

[1421] TGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTT

[1422] CGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGC

[1423] GCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATC

[1424] GCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGG

[1425] CGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGG

[1426] ACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGT TGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTT

[1427] GTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTT

[1428] TGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGG

[1429] GATTTTGGTCATGCATTCTAGGTACTAAAACAATTCATCCAGTAAAATATAATA

[1430] TTTTATTTTCTCCCAATCAGGCTTGATCCCCAGTAAGTCAAAAAATAGCTCGAC

[1431] ATACTGTTCTTCCCCGATATCCTCCCTGATCGACCGGACGCAGAAGGCAATG

[1432] TCATACCACTTGTCCGCCCTGCCGCTTCTCCCAAGATCAATAAAGCCACTTAC

[1433] TTTGCCATCTTTCACAAAGATGTTGCTGTCTCCCAGGTCGCCGTGGGAAAAG

[1434] ACAAGTTCCTCTTCGGGCTTTTCCGTCTTTAAAAAATCATACAGCTCGCGCGG

[1435] ATCTTTAAATGGAGTGTCTTCTTCCCAGTTTTCGCAATCCACATCGGCCAGAT

[1436] CGTTATTCAGTAAGTAATCCAATTCGGCTAAGCGGCTGTCTAAGCTATTCGTA

[1437] TAGGGACAATCCGATATGTCGATGGAGTGAAAGAGCCTGATGCACTCCGCAT

[1438] ACAGCTCGATAATCTTTTCAGGGCTTTGTTCATCTTCATACTCTTCCGAGCAAA

[1439] GGACGCCATCGGCCTCACTCATGAGCAGATTGCTCCAGCCATCATGCCGTTC

[1440] AAAGTGCAGGACCTTTGGAACAGGCAGCTTTCCTTCCAGCCATAGCATCATG

[1441] TCCTTTTCCCGTTCCACATCATAGGTGGTCCCTTTATACCGGCTGTCCGTCAT

[1442] TTTTAAATATAGGTTTTCATTTTCTCCCACCAGCTTATATACCTTAGCAGGAGA

[1443] CATTCCTTCCGTATCTTTTACGCAGCGGTATTTTTCGATCAGTTTTTTCAATTC

[1444] CGGTGATATTCTCATTTTAGCCATTTATTATTTCCTTCCTCTTTTCTACAGTATT

[1445] TAAAGATACCCCAAGAAGCTAATTATAACAAGACGAACTCCAATTCACTGTTC

[1446] CTTGCATTCTAAAACCTTAAATACCAGAAAACAGCTTTTTCAAAGTTGTTTTCA

[1447] AAGTTGGCGTATAACATAGTATCGACGGAGCCGATTTTGAAACCGCGGTGAT

[1448] CACAGGCAGCAACGCTCTGTCATCGTTACAATCAACATGCTACCCTCCGCGA

[1449] GATCATCCGTGTTTCAAACCCGGCAGCTTAGTTGCCGTTCTTCCGAATAGCAT

[1450] CGGTAACATGAGCAAAGTCTGCCGCCTTACAACGGCTCTCCCGCTGACGCC

[1451] GTCCCGGACTGATGGGCTGCCTGTATCGAGTGGTGATTTTGTGCCGAGCTGC

[1452] CGGTCGGGGAGCTGTTGGCTGGCTGGTGGCAGGATATATTGTGGTGTAAAC

[1453] AAATTGACGCTTAGACAACTTAATAACACATTGCGGACGTTTTTAATGTACTGA

[1454] ATTAACGCCGAATTGAATTCGAGCTCGGTACCGGGGGATCTGGATTTTAGTA

[1455] CTGGATTTTGGTTTTAGGAATTAGAAATTTTATTGATAGAAGTATTTTACAAATA

[1456] CAAATACATACTAAGGGTTTCTTATATGCTCAACACATGAGCGAAACCCTATA

[1457] GGAACCCTAATTCCCTTATCTGGGAACTACTCACACATTATTATGGAGAAACT

[1458] CGAGCTTGTCGATCGACTCAAATCTCGGTGACGGGCAGGACCGGACGGGGC

[1459] GGTACCGGCAGGCTGAAGTCCAGCTGCCAGAAACCCACGTCATGCCAGTTC

[1460] CCGTGCTTGAAGCCGGCCGCCCGCAGCATGCCGCGGGGGGCATATCCGAG

[1461] CGCCTCGTGCATGCGCACGCTCGGGTCGTTGGGCAGCCCGATGACAGCGAC

[1462] CACGCTCTTGAAGCCCTGTGCCTCCAGGGACTTCAGCAGGTGGGTGTAGAG

[1463] CGTGGAGCCCAGTCCCGTCCGCTGGTGGCGGGGGGAGACGTACACGGTCG ACTCGGCCGTCCAGTCGTAGGCGTTGCGTGCCTTCCAGGGGCCCGCGTAGG CGATGCCGGCGACCTCGCCGTCCACCTCGGCGACGAGCCAGGGATAGCGC TCCCGCAGACGGACGAGGTCGTCCGTCCACTCCTGCGGTTCCTGCGGCTCG GTACGGAAGTTGACCGTGCTTGTCTCGATGTAGTGGTTGACGATGGTGCAGA CCGCCGGCATGTCCGCCTCGGTGGCACGGCGGATGTCGGCCGGGCGTCGT TCTGGGCTCATGGTAGACGATCTGCGAAAGCTCGAGAGAGATAGATTTGTAG AGAGAGACTGGTGATTTCAGCGTGTCCTCTCCAAATGAAATGAACTTCCTTAT ATAGAGGAAGGGTCTTGCGAAGGATAGTGGGATTGTGCGTCATCCCTTACGT CAGTGGAGATATCACATCAATCCACTTGCTTTGAAGACGTGGTTGGAACGTCT TCTTTTTCCACGATGCTCCTCGTGGGTGGGGGTCCATCTTTGGGACCACTGT CGGCAGAGGCATCTTGAACGATAGCCTTTCCTTTATCGCAATGATGGCATTTG TAGGTGCCACCTTCCTTTTCTACTGTCCTTTTGATGAAGTGACAGATAGCTGG GCAATGGAATCCGAGGAGGTTTCCCGATATTACCCTTTGTTGAAAAGTCTCAA TAGCCCTTTGGTCTTCTGAGACTGTATCTTTGATATTCTTGGAGTAGACGAGA GTGTCGTGCTCCACCATGTTCACATCAATCCACTTGCTTTGAAGACGTGGTTG GAACGTCTTCTTTTTCCACGATGCTCCTCGTGGGTGGGGGTCCATCTTTGGG ACCACTGTCGGCAGAGGCATCTTGAACGATAGCCTTTCCTTTATCGCAATGAT GGCATTTGTAGGTGCCACCTTCCTTTTCTACTGTCCTTTTGATGAAGTGACAG ATAGCTGGGCAATGGAATCCGAGGAGGTTTCCCGATATTACCCTTTGTTGAAA AGTCTCAATAGCCCTTTGGTCTTCTGAGACTGTATCTTTGATATTCTTGGAGTA GACGAGAGTGTCGTGCTCCACCATGTTGGCGTACCCGGGGATCCTCTAGAG

[1464] TCGACCTGCAGGCATGCAAGCT

[1465] Prolyl 4-hydroxylase expression vector P4Hb_1 ;

[1466] SEQ ID NO: 30:

[1467] CAACTTTGTATAGAAAAGTTGGCCAACATGGTGGAGCACGACACTCTCGTCT ACTCCAAGAATATCAAAGATACAGTCTCAGAAGACCAAAGGGCTATTGAGACT TTTCAACAAAGGGTAATATCGGGAAACCTCCTCGGATTCCATTGCCCAGCTAT CTGTCACTTCATCAAAAGGACAGTAGAAAAGGAAGGTGGCACCTACAAATGC CATCATTGCGATAAAGGAAAGGCTATCGTTCAAGATGCCTCTGCCGACAGTG GTCCCAAAGATGGACCCCCACCCACGAGGAGCATCGTGGAAAAAGAAGACG TTCCAACCACGTCTTCAAAGCAAGTGGATTGATGTGAACATGGTGGAGCACG ACACTCTCGTCTACTCCAAGAATATCAAAGATACAGTCTCAGAAGACCAAAGG GCTATTGAGACTTTTCAACAAAGGGTAATATCGGGAAACCTCCTCGGATTCCA TTGCCCAGCTATCTGTCACTTCATCAAAAGGACAGTAGAAAAGGAAGGTGGC ACCTACAAATGCCATCATTGCGATAAAGGAAAGGCTATCGTTCAAGATGCCTC TGCCGACAGTGGTCCCAAAGATGGACCCCCACCCACGAGGAGCATCGTGGA AAAAGAAGACGTTCCAACCACGTCTTCAAAGCAAGTGGATTGATGTGATATCT CCACTGACGTAAGGGATGACGCACAATCCCACTATCCTTCGCAAGACCCTTC

[1468] CTCTATATAAGGAAGTTCATTTCATTTGGAGAGGACACGCTGACAAGTTTGTA

[1469] CAAAAAAGCAGGCTGCCACCATGAGAGTACAAAGAAGGAGGGCTCCAGATC

[1470] CTCCAGGTCAGTCTGAGGTTGGGACAGGAGGAAGGGCTAGAGGAGGAAGAA

[1471] CTTATAAATCTAGGACCGGAGGTGCTAGGTCAAGAGGACTTCTTAGCTGGAG

[1472] GAGACAGCTTAAGAGGAGGGTTTGCGCTGATATGCTTAGAAGGGCACTTCTT

[1473] TGCTTGGCTCTTACTGCTCTTTTTCGTGCTGGAGCTGGAGCTCCAGATGAGG

[1474] AGGATCATGTTTTGGTTCTTCATAAGGGTAATTTCGATGAAGCTCTTGCAGCT

[1475] CATAAGTATTTGCTTGTTGAGTTTTATGCTCCATGGTGTGGGCACTGTAAGGC

[1476] ACTTGCTCCAGAGTACGCTAAAGCTGCTGGCAAACTTAAGGCCGAGGGTTCA

[1477] GAAATTAGACTTGCAAAGGTGGATGCTACAGAGGAATCAGATTTGGCTCAAC

[1478] AGTATGGTGTTAGGGGTTATCCTACTATCAAGTTTTTTAAAAATGGTGATACAG

[1479] CTTCACCAAAGGAATATACAGCTGGAAGGGAAGCTGATGATATTGTTAATTGG

[1480] TTGAAGAAAAGAACTGGTCCAGCTGCTTCTACACTTTCAGATGGAGCTGCTG

[1481] CTGAAGCTCTTGTTGAAAGTTCCGAGGTTGCTGTTATTGGCTTTTTTAAGGAT

[1482] ATGGAATCCGATTCAGCTAAGCAGTTTTTGTTGGCAGCTGAAGCTATTGATGA

[1483] TATTCCTTTTGGAATTACTTCTAATTCTGACGTTTTCTCTAAATACCAATTGGAT

[1484] AAAGACGGTGTGGTTCTTTTCAAGAAGTTTGACGAAGGAAGAAACAATTTTGA

[1485] GGGTGAGGTTACCAAGGAGAAGCTTCTTGATTTTATCAAACATAACCAACTTC

[1486] CACTTGTTATTGAATTTACTGAACAAACTGCACCTAAGATTTTTGGTGGAGAAA

[1487] TTAAAACTCATATTCTCCTTTTCCTCCCTAAGAGCGTTAGCGATTATGAGGGTA

[1488] AGCTGTCAAATTTTAAGAAGGCTGCAGAATCATTTAAGGGTAAGATTTTGTTC

[1489] ATCTTTATCGATTCTGATCATACTGATAATCAAAGGATTTTGGAGTTTTTCGGT

[1490] TTGAAGAAGGAAGAGTGTCCTGCTGTTAGACTTATTACTTTGGAAGAGGAGAT

[1491] GACAAAATATAAACCTGAGTCTGATGAACTTACAGCAGAGAAAATTACTGAAT

[1492] TTTGCCACAGGTTTCTTGAAGGAAAGATTAAGCCTCATTTGATGAGTCAAGAA

[1493] TTGCCTGACGATTGGGATAAACAACCTGTTAAGGTTTTGGTTGGTAAAAACTT

[1494] CGAGGAGGTTGCTTTTGATGAGAAGAAGAACGTGTTTGTTGAATTTTATGCTC

[1495] CTTGGTGCGGACATTGTAAGCAGCTTGCTCCAATCTGGGATAAGTTGGGAGA

[1496] GACCTATAAAGATCATGAGAATATTGTTATTGCTAAAATGGATTCTACTGCAAA

[1497] CGAGGTTGAGGCAGTTAAGGTTCATAGTTTCCCTACTCTCAAGTTTTTTCCTG

[1498] CTTCTGCTGATAGAACTGTTATTGATTATAATGGTGAAAGAACACTCGATGGT

[1499] TTTAAGAAATTCCTTGAATCAGGAGGACAAGATGGTGCTGGTGACGATGATG

[1500] ATCTTGAAGATCTCGAAGAAGCTGAAGAGCCAGATTTGGAAGAAGATGATGA

[1501] TCAGAAGGCTGTTAAGGATGAGTTGGCATTTGTGTATTAAACCCAGCTTTCTT

[1502] GTACAAAGTGGTAGCTCGAATTTCCCCGATCGTTCAAACATTTGGCAATAAAG

[1503] TTTCTTAAGATTGAATCCTGTTGCCGGTCTTGCGATGATTATCATATAATTTCT

[1504] GTTGAATTACGTTAAGCATGTAATAATTAACATGTAATGCATGACGTTATTTAT GAGATGGGTTTTTATGATTAGAGTCCCGCAATTATACATTTAATACGCGATAG

[1505] AAAACAAAATATAGCGCGCAAACTAGGATAAATTATCGCGCGCGGTGTCATCT

[1506] ATGTTACTAGATCGGGAAAGCTTTTAAACTATCAGTGTTTGACAGGATATATTG

[1507] GCGGGTAAACCTAAGAGAAAAGAGCGTTTATTAGAATAATCGGATATTTAAAA

[1508] GGGCGTGAAAAGGTTTATCCGTTCGTCCATTTGTATGTGCATGCCAACCACA

[1509] GGGTTCCCCTCGGGATCAAAGTACTTTGATCCAACCCCTCCGCTGCTATAGT

[1510] GCAGTCGGCTTCTGACGTTCAGTGCAGCCGTCTTCTGAAAACGACATGTCGC

[1511] ACAAGTCCTAAGTTACGCGACAGGCTGCCGCCCTGCCCTTTTCCTGGCGTTT

[1512] TCTTGTCGCGTGTTTTAGTCGCATAAAGTAGAATACTTGCGACTAGAACCGGA

[1513] GACATTACGCCATGAACAAGAGCGCCGCCGCTGGCCTGCTGGGCTATGCCC

[1514] GCGTCAGCACCGACGACCAGGACTTGACCAACCAACGGGCCGAACTGCACG

[1515] CGGCCGGCTGCACCAAGCTGTTTTCCGAGAAGATCACCGGCACCAGGCGCG

[1516] ACCGCCCGGAGCTGGCCAGGATGCTTGACCACCTACGCCCTGGCGACGTTG

[1517] TGACAGTGACCAGGCTAGACCGCCTGGCCCGCAGCACCCGCGACCTACTGG

[1518] ACATTGCCGAGCGCATCCAGGAGGCCGGCGCGGGCCTGCGTAGCCTGGCA

[1519] GAGCCGTGGGCCGACACCACCACGCCGGCCGGCCGCATGGTGTTGACCGT

[1520] GTTCGCCGGCATTGCCGAGTTCGAGCGTTCCCTAATCATCGACCGCACCCG

[1521] GAGCGGGCGCGAGGCCGCCAAGGCCCGAGGCGTGAAGTTTGGCCCCCGCC

[1522] CTACCCTCACCCCGGCACAGATCGCGCACGCCCGCGAGCTGATCGACCAGG

[1523] AAGGCCGCACCGTGAAAGAGGCGGCTGCACTGCTTGGCGTGCATCGCTCGA

[1524] CCCTGTACCGCGCACTTGAGCGCAGCGAGGAAGTGACGCCCACCGAGGCCA

[1525] GGCGGCGCGGTGCCTTCCGTGAGGACGCATTGACCGAGGCCGACGCCCTG

[1526] GCGGCCGCCGAGAATGAACGCCAAGAGGAACAAGCATGAAACCGCACCAGG

[1527] ACGGCCAGGACGAACCGTTTTTCATTACCGAAGAGATCGAGGCGGAGATGAT

[1528] CGCGGCCGGGTACGTGTTCGAGCCGCCCGCGCACGTCTCAACCGTGCGGC

[1529] TGCATGAAATCCTGGCCGGTTTGTCTGATGCCAAGCTGGCGGCCTGGCCGG

[1530] CCAGCTTGGCCGCTGAAGAAACCGAGCGCCGCCGTCTAAAAAGGTGATGTG

[1531] TATTTGAGTAAAACAGCTTGCGTCATGCGGTCGCTGCGTATATGATGCGATGA

[1532] GTAAATAAACAAATACGCAAGGGGAACGCATGAAGGTTATCGCTGTACTTAAC

[1533] CAGAAAGGCGGGTCAGGCAAGACGACCATCGCAACCCATCTAGCCCGCGCC

[1534] CTGCAACTCGCCGGGGCCGATGTTCTGTTAGTCGATTCCGATCCCCAGGGCA

[1535] GTGCCCGCGATTGGGCGGCCGTGCGGGAAGATCAACCGCTAACCGTTGTCG

[1536] GCATCGACCGCCCGACGATTGACCGCGACGTGAAGGCCATCGGCCGGCGC

[1537] GACTTCGTAGTGATCGACGGAGCGCCCCAGGCGGCGGACTTGGCTGTGTCC

[1538] GCGATCAAGGCAGCCGACTTCGTGCTGATTCCGGTGCAGCCAAGCCCTTAC

[1539] GACATATGGGCCACCGCCGACCTGGTGGAGCTGGTTAAGCAGCGCATTGAG

[1540] GTCACGGATGGAAGGCTACAAGCGGCCTTTGTCGTGTCGCGGGCGATCAAA

[1541] GGCACGCGCATCGGCGGTGAGGTTGCCGAGGCGCTGGCCGGGTACGAGCT GCCCATTCTTGAGTCCCGTATCACGCAGCGCGTGAGCTACCCAGGCACTGC

[1542] CGCCGCCGGCACAACCGTTCTTGAATCAGAACCCGAGGGCGACGCTGCCCG

[1543] CGAGGTCCAGGCGCTGGCCGCTGAAATTAAATCAAAACTCATTTGAGTTAAT

[1544] GAGGTAAAGAGAAAATGAGCAAAAGCACAAACACGCTAAGTGCCGGCCGTC

[1545] CGAGCGCACGCAGCAGCAAGGCTGCAACGTTGGCCAGCCTGGCAGACACG

[1546] CCAGCCATGAAGCGGGTCAACTTTCAGTTGCCGGCGGAGGATCACACCAAG

[1547] CTGAAGATGTACGCGGTACGCCAAGGCAAGACCATTACCGAGCTGCTATCTG

[1548] AATACATCGCGCAGCTACCAGAGTAAATGAGCAAATGAATAAATGAGTAGATG

[1549] AATTTTAGCGGCTAAAGGAGGCGGCATGGAAAATCAAGAACAACCAGGCACC

[1550] GACGCCGTGGAATGCCCCATGTGTGGAGGAACGGGCGGTTGGCCAGGCGT

[1551] AAGCGGCTGGGTTGCCTGCCGGCCCTGCAATGGCACTGGAACCCCCAAGCC

[1552] CGAGGAATCGGCGTGAGCGGTCGCAAACCATCCGGCCCGGTACAAATCGGC

[1553] GCGGCGCTGGGTGATGACCTGGTGGAGAAGTTGAAGGCCGCGCAGGCCGC

[1554] CCAGCGGCAACGCATCGAGGCAGAAGCACGCCCCGGTGAATCGTGGCAAG

[1555] CGGCCGCTGATCGAATCCGCAAAGAATCCCGGCAACCGCCGGCAGCCGGTG

[1556] CGCCGTCGATTAGGAAGCCGCCCAAGGGCGACGAGCAACCAGATTTTTTCGT

[1557] TCCGATGCTCTATGACGTGGGCACCCGCGATAGTCGCAGCATCATGGACGT

[1558] GGCCGTTTTCCGTCTGTCGAAGCGTGACCGACGAGCTGGCGAGGTGATCCG

[1559] CTACGAGCTTCCAGACGGGCACGTAGAGGTTTCCGCAGGGCCGGCCGGCAT

[1560] GGCCAGTGTGTGGGATTACGACCTGGTACTGATGGCGGTTTCCCATCTAACC

[1561] GAATCCATGAACCGATACCGGGAAGGGAAGGGAGACAAGCCCGGCCGCGT

[1562] GTTCCGTCCACACGTTGCGGACGTACTCAAGTTCTGCCGGCGAGCCGATGG

[1563] CGGAAAGCAGAAAGACGACCTGGTAGAAACCTGCATTCGGTTAAACACCACG

[1564] CACGTTGCCATGCAGCGTACGAAGAAGGCCAAGAACGGCCGCCTGGTGACG

[1565] GTATCCGAGGGTGAAGCCTTGATTAGCCGCTACAAGATCGTAAAGAGCGAAA

[1566] CCGGGCGGCCGGAGTACATCGAGATCGAGCTAGCTGATTGGATGTACCGCG

[1567] AGATCACAGAAGGCAAGAACCCGGACGTGCTGACGGTTCACCCCGATTACTT

[1568] TTTGATCGATCCCGGCATCGGCCGTTTTCTCTACCGCCTGGCACGCCGCGCC

[1569] GCAGGCAAGGCAGAAGCCAGATGGTTGTTCAAGACGATCTACGAACGCAGT

[1570] GGCAGCGCCGGAGAGTTCAAGAAGTTCTGTTTCACCGTGCGCAAGCTGATC

[1571] GGGTCAAATGACCTGCCGGAGTACGATTTGAAGGAGGAGGCGGGGCAGGCT

[1572] GGCCCGATCCTAGTCATGCGCTACCGCAACCTGATCGAGGGCGAAGCATCC

[1573] GCCGGTTCCTAATGTACGGAGCAGATGCTAGGGCAAATTGCCCTAGCAGGG

[1574] GAAAAAGGTCGAAAAGGTCTCTTTCCTGTGGATAGCACGTACATTGGGAACC

[1575] CAAAGCCGTACATTGGGAACCGGAACCCGTACATTGGGAACCCAAAGCCGTA

[1576] CATTGGGAACCGGTCACACATGTAAGTGACTGATATAAAAGAGAAAAAAGGC

[1577] GATTTTTCCGCCTAAAACTCTTTAAAACTTATTAAAACTCTTAAAACCCGCCTG

[1578] GCCTGTGCATAACTGTCTGGCCAGCGCACAGCCGAAGAGCTGCAAAAAGCG CCTACCCTTCGGTCGCTGCGCTCCCTACGCCCCGCCGCTTCGCGTCGGCCT

[1579] ATCGCGGCCGCTGGCCGCTCAAAAATGGCTGGCCTACGGCCAGGCAATCTA

[1580] CCAGGGCGCGGACAAGCCGCGCCGTCGCCACTCGACCGCCGGCGCCCACA

[1581] TCAAGGCACCCTGCCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACA

[1582] CATGCAGCTCCCGGAGACGGTCACAGCTTGTCTGTAAGCGGATGCCGGGAG

[1583] CAGACAAGCCCGTCAGGGCGCGTCAGCGGGTGTTGGCGGGTGTCGGGGCG

[1584] CAGCCATGACCCAGTCACGTAGCGATAGCGGAGTGTATACTGGCTTAACTAT

[1585] GCGGCATCAGAGCAGATTGTACTGAGAGTGCACCATATGCGGTGTGAAATAC

[1586] CGCACAGATGCGTAAGGAGAAAATACCGCATCAGGCGCTCTTCCGCTTCCTC

[1587] GCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGC

[1588] TCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGA

[1589] AAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCC

[1590] GCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAA

[1591] ATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCA

[1592] GGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCC

[1593] GCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCT

[1594] CATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGC

[1595] TGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCG

[1596] GTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGC

[1597] AGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTAC

[1598] AGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTT

[1599] GGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCT

[1600] CTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAA

[1601] GCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTT

[1602] CTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGT

[1603] CATGCATTCTAGGTACTAAAACAATTCATCCAGTAAAATATAATATTTTATTTTC

[1604] TCCCAATCAGGCTTGATCCCCAGTAAGTCAAAAAATAGCTCGACATACTGTTC

[1605] TTCCCCGATATCCTCCCTGATCGACCGGACGCAGAAGGCAATGTCATACCAC

[1606] TTGTCCGCCCTGCCGCTTCTCCCAAGATCAATAAAGCCACTTACTTTGCCATC

[1607] TTTCACAAAGATGTTGCTGTCTCCCAGGTCGCCGTGGGAAAAGACAAGTTCC

[1608] TCTTCGGGCTTTTCCGTCTTTAAAAAATCATACAGCTCGCGCGGATCTTTAAA

[1609] TGGAGTGTCTTCTTCCCAGTTTTCGCAATCCACATCGGCCAGATCGTTATTCA

[1610] GTAAGTAATCCAATTCGGCTAAGCGGCTGTCTAAGCTATTCGTATAGGGACAA

[1611] TCCGATATGTCGATGGAGTGAAAGAGCCTGATGCACTCCGCATACAGCTCGA

[1612] TAATCTTTTCAGGGCTTTGTTCATCTTCATACTCTTCCGAGCAAAGGACGCCA

[1613] TCGGCCTCACTCATGAGCAGATTGCTCCAGCCATCATGCCGTTCAAAGTGCA

[1614] GGACCTTTGGAACAGGCAGCTTTCCTTCCAGCCATAGCATCATGTCCTTTTCC

[1615] CGTTCCACATCATAGGTGGTCCCTTTATACCGGCTGTCCGTCATTTTTAAATA TAGGTTTTCATTTTCTCCCACCAGCTTATATACCTTAGCAGGAGACATTCCTTC

[1616] CGTATCTTTTACGCAGCGGTATTTTTCGATCAGTTTTTTCAATTCCGGTGATAT

[1617] TCTCATTTTAGCCATTTATTATTTCCTTCCTCTTTTCTACAGTATTTAAAGATAC

[1618] CCCAAGAAGCTAATTATAACAAGACGAACTCCAATTCACTGTTCCTTGCATTC

[1619] TAAAACCTTAAATACCAGAAAACAGCTTTTTCAAAGTTGTTTTCAAAGTTGGCG

[1620] TATAACATAGTATCGACGGAGCCGATTTTGAAACCGCGGTGATCACAGGCAG

[1621] CAACGCTCTGTCATCGTTACAATCAACATGCTACCCTCCGCGAGATCATCCGT

[1622] GTTTCAAACCCGGCAGCTTAGTTGCCGTTCTTCCGAATAGCATCGGTAACATG

[1623] AGCAAAGTCTGCCGCCTTACAACGGCTCTCCCGCTGACGCCGTCCCGGACT

[1624] GATGGGCTGCCTGTATCGAGTGGTGATTTTGTGCCGAGCTGCCGGTCGGGG

[1625] AGCTGTTGGCTGGCTGGTGGCAGGATATATTGTGGTGTAAACAAATTGACGC TTAGACAACTTAATAACACATTGCGGACGTTTTTAATGTACTGAATTAACGCCG

[1626] AATTGAATTCGAGCTCGGTACCGGGGGATCTGGATTTTAGTACTGGATTTTGG

[1627] TTTTAGGAATTAGAAATTTTATTGATAGAAGTATTTTACAAATACAAATACATAC

[1628] TAAGGGTTTCTTATATGCTCAACACATGAGCGAAACCCTATAGGAACCCTAAT

[1629] TCCCTTATCTGGGAACTACTCACACATTATTATGGAGAAACTCGAGCTTGTCG

[1630] ATCGACTCAAATCTCGGTGACGGGCAGGACCGGACGGGGCGGTACCGGCA

[1631] GGCTGAAGTCCAGCTGCCAGAAACCCACGTCATGCCAGTTCCCGTGCTTGAA

[1632] GCCGGCCGCCCGCAGCATGCCGCGGGGGGCATATCCGAGCGCCTCGTGCA

[1633] TGCGCACGCTCGGGTCGTTGGGCAGCCCGATGACAGCGACCACGCTCTTGA

[1634] AGCCCTGTGCCTCCAGGGACTTCAGCAGGTGGGTGTAGAGCGTGGAGCCCA

[1635] GTCCCGTCCGCTGGTGGCGGGGGGAGACGTACACGGTCGACTCGGCCGTC

[1636] CAGTCGTAGGCGTTGCGTGCCTTCCAGGGGCCCGCGTAGGCGATGCCGGC

[1637] GACCTCGCCGTCCACCTCGGCGACGAGCCAGGGATAGCGCTCCCGCAGAC

[1638] GGACGAGGTCGTCCGTCCACTCCTGCGGTTCCTGCGGCTCGGTACGGAAGT

[1639] TGACCGTGCTTGTCTCGATGTAGTGGTTGACGATGGTGCAGACCGCCGGCAT

[1640] GTCCGCCTCGGTGGCACGGCGGATGTCGGCCGGGCGTCGTTCTGGGCTCAT

[1641] GGTAGACGATCTGCGAAAGCTCGAGAGAGATAGATTTGTAGAGAGAGACTGG

[1642] TGATTTCAGCGTGTCCTCTCCAAATGAAATGAACTTCCTTATATAGAGGAAGG

[1643] GTCTTGCGAAGGATAGTGGGATTGTGCGTCATCCCTTACGTCAGTGGAGATA

[1644] TCACATCAATCCACTTGCTTTGAAGACGTGGTTGGAACGTCTTCTTTTTCCAC

[1645] GATGCTCCTCGTGGGTGGGGGTCCATCTTTGGGACCACTGTCGGCAGAGGC

[1646] ATCTTGAACGATAGCCTTTCCTTTATCGCAATGATGGCATTTGTAGGTGCCAC

[1647] CTTCCTTTTCTACTGTCCTTTTGATGAAGTGACAGATAGCTGGGCAATGGAAT

[1648] CCGAGGAGGTTTCCCGATATTACCCTTTGTTGAAAAGTCTCAATAGCCCTTTG

[1649] GTCTTCTGAGACTGTATCTTTGATATTCTTGGAGTAGACGAGAGTGTCGTGCT

[1650] CCACCATGTTCACATCAATCCACTTGCTTTGAAGACGTGGTTGGAACGTCTTC

[1651] TTTTTCCACGATGCTCCTCGTGGGTGGGGGTCCATCTTTGGGACCACTGTCG GCAGAGGCATCTTGAACGATAGCCTTTCCTTTATCGCAATGATGGCATTTGTA

[1652] GGTGCCACCTTCCTTTTCTACTGTCCTTTTGATGAAGTGACAGATAGCTGGGC

[1653] AATGGAATCCGAGGAGGTTTCCCGATATTACCCTTTGTTGAAAAGTCTCAATA

[1654] GCCCTTTGGTCTTCTGAGACTGTATCTTTGATATTCTTGGAGTAGACGAGAGT

[1655] GTCGTGCTCCACCATGTTGGCGTACCCGGGGATCCTCTAGAGTCGACCTGCA

[1656] GGCATGCAAGCT

[1657] Bovine Collagen alpha-1 (I) Expression vector 7;

[1658] SEQ ID NO: 31 :

[1659] AGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCG

[1660] CCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAA

[1661] CCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTG

[1662] CGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCC

[1663] CTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTC

[1664] GGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCA

[1665] GCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTA

[1666] AGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAG

[1667] CGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGG

[1668] CTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCT

[1669] TCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAG

[1670] CGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTC

[1671] AAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAAC

[1672] TCACGTTAAGGGATTTTGGTCATGCATTCTAGGTACTAAAACAATTCATCCAG

[1673] TAAAATATAATATTTTATTTTCTCCCAATCAGGCTTGATCCCCAGTAAGTCAAA

[1674] AAATAGCTCGACATACTGTTCTTCCCCGATATCCTCCCTGATCGACCGGACG

[1675] CAGAAGGCAATGTCATACCACTTGTCCGCCCTGCCGCTTCTCCCAAGATCAA

[1676] TAAAGCCACTTACTTTGCCATCTTTCACAAAGATGTTGCTGTCTCCCAGGTCG

[1677] CCGTGGGAAAAGACAAGTTCCTCTTCGGGCTTTTCCGTCTTTAAAAAATCATA

[1678] CAGCTCGCGCGGATCTTTAAATGGAGTGTCTTCTTCCCAGTTTTCGCAATCCA

[1679] CATCGGCCAGATCGTTATTCAGTAAGTAATCCAATTCGGCTAAGCGGCTGTCT

[1680] AAGCTATTCGTATAGGGACAATCCGATATGTCGATGGAGTGAAAGAGCCTGA

[1681] TGCACTCCGCATACAGCTCGATAATCTTTTCAGGGCTTTGTTCATCTTCATACT

[1682] CTTCCGAGCAAAGGACGCCATCGGCCTCACTCATGAGCAGATTGCTCCAGCC

[1683] ATCATGCCGTTCAAAGTGCAGGACCTTTGGAACAGGCAGCTTTCCTTCCAGC

[1684] CATAGCATCATGTCCTTTTCCCGTTCCACATCATAGGTGGTCCCTTTATACCG

[1685] GCTGTCCGTCATTTTTAAATATAGGTTTTCATTTTCTCCCACCAGCTTATATAC

[1686] CTTAGCAGGAGACATTCCTTCCGTATCTTTTACGCAGCGGTATTTTTCGATCA

[1687] GTTTTTTCAATTCCGGTGATATTCTCATTTTAGCCATTTATTATTTCCTTCCTCT TTTCTACAGTATTTAAAGATACCCCAAGAAGCTAATTATAACAAGACGAACTCC AATTCACTGTTCCTTGCATTCTAAAACCTTAAATACCAGAAAACAGCTTTTTCA AAGTTGTTTTCAAAGTTGGCGTATAACATAGTATCGACGGAGCCGATTTTGAA ACCGCGGTGATCACAGGCAGCAACGCTCTGTCATCGTTACAATCAACATGCT ACCCTCCGCGAGATCATCCGTGTTTCAAACCCGGCAGCTTAGTTGCCGTTCT

[1688] TCCGAATAGCATCGGTAACATGAGCAAAGTCTGCCGCCTTACAACGGCTCTC CCGCTGACGCCGTCCCGGACTGATGGGCTGCCTGTATCGAGTGGTGATTTT

[1689] GTGCCGAGCTGCCGGTCGGGGAGCTGTTGGCTGGCTGGTGGCAGGATATAT TGTGGTGTAAACAAATTGACGCTTAGACAACTTAATAACACATTGCGGACGTT

[1690] TTTAATGTACTGAATTAACGCCGAATTGAATTCGAGCTCGGTGGAGGATCTGG ATTTTAGTACTGGATTTTGGTTTTAGGAATTAGAAATTTTATTGATAGAAGTATT TTACAAATACAAATACATACTAAGGGTTTCTTATATGCTCAACACATGAGCGAA ACCCTATAGGAACCCTAATTCCCTTATCTGGGAACTACTCACACATTATTATG GAGAAACTATTCACTCTCCTTCTCAAACGTTTCTGTTCCCTCTGGACAGCCCC

[1691] GACTGACGTTGGACTCAACCTCAATGGGCGCTAATTGTAATAGCTCCTGTTTG CTTCTTATGGCCATTTCGCAAAGGTGTTGCAGTGTTCTGCTACCACCACTCAC TGTAATTGACACACCTCTGAAACGGATGGAGTAAGTGCAACCAATTTGATCAA ATCCAAAGAAACGACTAGCAGCGTAATTTACTGAGTCACCAGTCCAGGAACC TAGAACTCGATGGAGAGAGGCCTCTGTCCGATCATAGCGTAGGTAGCGTTTG

[1692] AAAACTACCTTGCCAAATCCCCAGGACTCTTTAAATCCCAGTGGGTTGTCCTG GTTGGAGTTTGTTTCGTCATTGTGTAAACGCCATTCTGTCCAAGATGGGCTCT

[1693] CATCGGGTAACTTAAATGGGCTGGTAGTGCCGCCAGAACCACCGTCCCATCT CTCTGAATTGGCCTGCTCTCTTGCATCATTACCCTGGATTGCTCTCTCCATTG

[1694] TCCTCTCCAAATGAAATGAACTTCCTTATATAGAGGAAGGGTCTTGCGAAGGA TAGTGGGATTGTGCGTCATCCCTTACGTCAGTGGAGATGTCACATCAATCCA CTTGCTTTGTAGACGTGGTTGGAACCTCTTCTTTTTCCACGATGCTCCTCGTG

[1695] GGTGGGGGTCCATCTTTGGGACCACTGTCGGCAGAGAGATCTTGAATGATAG CCTTTCCTTTATCGCAATGATGGCATTTGTAGGAGCCACCTTCCTTTTCTACT

[1696] GTCCTTTCGATGAAGTGACAGATAGCTGGGCAATGGAATCCGAGGAGGTTTC CCGAAATTATCCTTTGTTGAAAAGTCTCAAACAATAGGCAACCGTGGACTTCT

[1697] TCACTAGCTCATCGAGATAGCAATGCCACTAGCTAATTTCTTACGTTGTATCTT

[1698] ATTTGTTTTACTTTGGGGCATGACATGGTTAAACCCCATCAAAGAGAAAGTGT TTCATCCACTAGTCAATATACGAATTCATTCGAATTTATCCCTGTAAATCCTAG

[1699] TTCTTAGGATGAACTGGTGTAATAAACAGCAAAAAAAAATAAAAAAAAATAAAT CTGGAATCATTCGACCACCTCAATAAACTAAAGCTACCAATTACCACAATATA GTC ATC C ATATC C ACTTAGATATAAAAGATAAAAGTAAAC AAATATTAAATTTC ATATGCACGCATAGGAAACTCATGATCTTATCTTTTAAATAGACATCTAGTTTT CTTAGGTTATAAATAGACATTTTGTCCTAGAACTTCTTCACTACTAAAACCTAG CCTCAAAAAGCAGGCTGCCACCATGCATCACCATCATCATCATGGATCTGAG

[1700] AATTTATATTTTCAAGGTCAATTGAGTTATGGATACGACGAAAAATCAACTGGA

[1701] ATATCAGTGCCAGGACCAATGGGTCCATCAGGACCTAGGGGTCTTCCAGGG

[1702] CCACCTGGCGCTCCTGGTCCTCAGGGATTTCAAGGTCCTCCAGGTGAACCTG

[1703] GAGAGCCTGGTGCTTCTGGTCCAATGGGACCAAGAGGCCCTCCTGGTCCAC

[1704] CTGGTAAAAATGGTGATGATGGTGAGGCTGGAAAGCCTGGTCGTCCTGGAG

[1705] AAAGAGGACCACCAGGACCACAGGGAGCTAGAGGTTTGCCAGGTACAGCCG

[1706] GTCTTCCAGGAATGAAAGGACACAGGGGTTTTTCTGGTCTTGATGGTGCTAA

[1707] GGGAGACGCAGGACCTGCTGGACCAAAGGGTGAACCTGGATCTCCTGGTGA

[1708] AAATGGTGCTCCTGGACAAATGGGACCTAGAGGATTGCCTGGCGAAAGAGG

[1709] AAGACCTGGTGCTCCTGGACCAGCTGGAGCTAGAGGAAATGATGGAGCTAC

[1710] TGGTGCTGCCGGACCACCAGGACCAACTGGACCAGCTGGACCACCTGGTTT

[1711] TCCAGGAGCAGTTGGGGCTAAGGGAGAAGGTGGACCACAAGGACCTAGAGG

[1712] ATCAGAAGGACCTCAAGGAGTTAGGGGAGAACCAGGTCCTCCTGGTCCTGC

[1713] TGGAGCTGCTGGACCCGCTGGTAACCCAGGAGCAGATGGACAACCTGGAGC

[1714] AAAAGGGGCTAATGGTGCTCCTGGTATTGCAGGTGCTCCTGGATTCCCTGGT

[1715] GCTAGAGGACCTTCAGGACCTCAGGGACCATCCGGTCCTCCAGGACCAAAG

[1716] GGAAACTCTGGTGAACCAGGGGCTCCTGGATCTAAGGGAGATACTGGAGCT

[1717] AAGGGTGAGCCAGGACCTACAGGAATTCAAGGACCACCAGGACCTGCAGGA

[1718] GAGGAAGGTAAGAGGGGTGCTAGAGGTGAACCTGGACCAGCTGGATTGCCA

[1719] GGCCCACCTGGAGAGAGGGGTGGTCCAGGTTCTAGAGGTTTTCCAGGAGCT

[1720] GATGGAGTTGCTGGTCCAAAGGGTCCTGCTGGTGAGAGAGGTGCACCTGGA

[1721] CCAGCAGGACCTAAAGGATCACCTGGAGAAGCTGGTCGTCCTGGTGAGGCT

[1722] GGTCTTCCAGGTGCTAAGGGCCTTACAGGATCTCCAGGATCACCAGGTCCAG

[1723] ATGGAAAGACTGGACCACCAGGTCCTGCAGGACAAGATGGAAGGCCTGGTC

[1724] CACCTGGACCACCTGGTGCTAGGGGACAAGCTGGTGTTATGGGATTCCCAG

[1725] GTCCAAAGGGTGCAGCTGGTGAGCCTGGTAAAGCTGGTGAAAGAGGAGTTC

[1726] CAGGACCTCCTGGTGCTGTTGGACCTGCTGGTAAAGATGGTGAAGCAGGTG

[1727] CTCAGGGTCCACCAGGTCCAGCCGGTCCAGCTGGTGAGAGGGGAGAGCAA

[1728] GGTCCTGCTGGATCTCCTGGATTTCAAGGTTTGCCTGGACCTGCTGGACCTC

[1729] CAGGAGAGGCTGGTAAACCTGGTGAGCAGGGTGTGCCTGGTGATCTTGGTG

[1730] CTCCTGGACCTAGTGGCGCAAGAGGCGAAAGGGGATTTCCAGGTGAAAGAG

[1731] GAGTTCAGGGACCACCAGGTCCAGCTGGTCCAAGAGGCGCTAATGGAGCTC

[1732] CAGGAAATGATGGAGCAAAGGGTGATGCTGGCGCTCCAGGAGCTCCTGGTT

[1733] CTCAAGGTGCACCTGGATTGCAAGGAATGCCAGGAGAGAGAGGTGCAGCTG

[1734] GTTTGCCGGGACCAAAGGGTGATAGAGGTGATGCTGGTCCTAAGGGAGCTG

[1735] ATGGAGCTCCAGGAAAGGATGGTGTGAGAGGATTGACTGGTCCAATTGGAC

[1736] CACCAGGTCCTGCTGGAGCACCAGGAGATAAAGGAGAAGCTGGTCCTTCAG GACCAGCCGGTCCAACTGGAGCTAGAGGAGCTCCAGGAGATAGAGGTGAAC CAGGACCACCAGGACCTGCTGGGTTTGCAGGTCCACCTGGTGCTGATGGAC AACCTGGTGCTAAAGGAGAACCAGGCGATGCTGGTGCTAAGGGAGATGCTG GCCCACCTGGTCCAGCTGGTCCTGCTGGGCCACCAGGTCCTATCGGAAATG TTGGTGCTCCAGGGCCTAAAGGAGCTAGAGGATCAGCAGGTCCCCCAGGTG CTACAGGTTTTCCAGGGGCTGCTGGAAGAGTGGGTCCCCCAGGTCCTTCTG GGAATGCAGGACCACCAGGTCCTCCAGGACCAGCTGGAAAAGAAGGAAGTA AGGGACCAAGGGGAGAAACTGGACCTGCAGGACGTCCAGGTGAAGTTGGTC CACCTGGACCTCCAGGTCCAGCTGGAGAGAAAGGAGCTCCAGGAGCTGATG GTCCAGCAGGTGCTCCTGGTACTCCAGGTCCACAAGGTATTGCTGGTCAAAG GGGAGTGGTTGGTCTCCCAGGTCAGAGAGGTGAAAGAGGTTTCCCAGGTTT GCCTGGACCATCTGGAGAACCAGGTAAGCAGGGCCCATCTGGAGCTTCTGG TGAGAGAGGGCCACCTGGACCTATGGGACCTCCTGGGCTTGCTGGACCTCC AGGGGAATCTGGTAGGGAGGGTGCTCCAGGTGCTGAAGGATCACCTGGTAG

[1737] AGATGGATCACCTGGAGCTAAAGGTGATAGAGGGGAAACAGGTCCAGCAGG TCCACCTGGCGCTCCTGGTGCTCCAGGAGCACCAGGGCCAGTTGGACCTGC TGGAAAGTCTGGAGATAGGGGAGAGACTGGACCAGCTGGACCTGCTGGACC AATCGGTCCTGTTGGCGCTAGAGGTCCTGCTGGACCACAAGGTCCTAGGGG AGATAAAGGTGAAACAGGTGAACAAGGAGATAGAGGAATTAAAGGTCATAGA GGGTTTTCTGGTTTGCAAGGACCTCCAGGACCTCCTGGATCTCCAGGAGAGC AAGGACCATCTGGTGCAAGCGGACCTGCTGGTCCTAGAGGACCTCCAGGTT CAGCTGGCTCACCTGGTAAGGATGGATTAAATGGTTTGCCTGGACCAATAGG TCCACCAGGACCTAGAGGAAGGACAGGTGATGCAGGACCAGCTGGTCCACC TGGTCCACCAGGACCACCAGGACCACCAGGTCCACCTTCTGGTGGATATGAT CTTAGTTTTCTTCCTCAACCTCCCCAGGAGAAAGCTCATGATGGTGGTAGATA TTATAGGGCTGCATTTGTTTATTAAACCCAGCTTTCTTGTACAAAGTGGTAGCT CGAATTTCCCCGATCGTTCAAACATTTGGCAATAAAGTTTCTTAAGATTGAATC CTGTTGCCGGTCTTGCGATGATTATCATATAATTTCTGTTGAATTACGTTAAGC

[1738] ATGTAATAATTAACATGTAATGCATGACGTTATTTATGAGATGGGTTTTTATGA TTAGAGTCCCGCAATTATACATTTAATACGCGATAGAAAACAAAATATAGCGC GCAAACTAGGATAAATTATCGCGCGCGGTGTCATCTATGTTACTAGATCGGG AAAGCTTTTAAACTATCAGTGTTTGACAGGATATATTGGCGGGTAAACCTAAG AGAAAAGAGCGTTTATTAGAATAATCGGATATTTAAAAGGGCGTGAAAAGGTT TATCCGTTCGTCCATTTGTATGTGCATGCCAACCACAGGGTTCCCCTCGGGA TCAAAGTACTTTGATCCAACCCCTCCGCTGCTATAGTGCAGTCGGCTTCTGAC GTTCAGTGCAGCCGTCTTCTGAAAACGACATGTCGCACAAGTCCTAAGTTAC GCGACAGGCTGCCGCCCTGCCCTTTTCCTGGCGTTTTCTTGTCGCGTGTTTT AGTCGCATAAAGTAGAATACTTGCGACTAGAACCGGAGACATTACGCCATGA ACAAGAGCGCCGCCGCTGGCCTGCTGGGCTATGCCCGCGTCAGCACCGAC

[1739] GACCAGGACTTGACCAACCAACGGGCCGAACTGCACGCGGCCGGCTGCACC

[1740] AAGCTGTTTTCCGAGAAGATCACCGGCACCAGGCGCGACCGCCCGGAGCTG

[1741] GCCAGGATGCTTGACCACCTACGCCCTGGCGACGTTGTGACAGTGACCAGG

[1742] CTAGACCGCCTGGCCCGCAGCACCCGCGACCTACTGGACATTGCCGAGCGC

[1743] ATCCAGGAGGCCGGCGCGGGCCTGCGTAGCCTGGCAGAGCCGTGGGCCGA

[1744] CACCACCACGCCGGCCGGCCGCATGGTGTTGACCGTGTTCGCCGGCATTGC

[1745] CGAGTTCGAGCGTTCCCTAATCATCGACCGCACCCGGAGCGGGCGCGAGGC

[1746] CGCCAAGGCCCGAGGCGTGAAGTTTGGCCCCCGCCCTACCCTCACCCCGGC

[1747] ACAGATCGCGCACGCCCGCGAGCTGATCGACCAGGAAGGCCGCACCGTGAA

[1748] AGAGGCGGCTGCACTGCTTGGCGTGCATCGCTCGACCCTGTACCGCGCACT

[1749] TGAGCGCAGCGAGGAAGTGACGCCCACCGAGGCCAGGCGGCGCGGTGCCT

[1750] TCCGTGAGGACGCATTGACCGAGGCCGACGCCCTGGCGGCCGCCGAGAAT

[1751] GAACGCCAAGAGGAACAAGCATGAAACCGCACCAGGACGGCCAGGACGAAC

[1752] CGTTTTTCATTACCGAAGAGATCGAGGCGGAGATGATCGCGGCCGGGTACGT

[1753] GTTCGAGCCGCCCGCGCACGTCTCAACCGTGCGGCTGCATGAAATCCTGGC

[1754] CGGTTTGTCTGATGCCAAGCTGGCGGCCTGGCCGGCCAGCTTGGCCGCTGA

[1755] AGAAACCGAGCGCCGCCGTCTAAAAAGGTGATGTGTATTTGAGTAAAACAGC

[1756] TTGCGTCATGCGGTCGCTGCGTATATGATGCGATGAGTAAATAAACAAATAC

[1757] GCAAGGGGAACGCATGAAGGTTATCGCTGTACTTAACCAGAAAGGCGGGTC

[1758] AGGCAAGACGACCATCGCAACCCATCTAGCCCGCGCCCTGCAACTCGCCGG

[1759] GGCCGATGTTCTGTTAGTCGATTCCGATCCCCAGGGCAGTGCCCGCGATTG

[1760] GGCGGCCGTGCGGGAAGATCAACCGCTAACCGTTGTCGGCATCGACCGCCC

[1761] GACGATTGACCGCGACGTGAAGGCCATCGGCCGGCGCGACTTCGTAGTGAT

[1762] CGACGGAGCGCCCCAGGCGGCGGACTTGGCTGTGTCCGCGATCAAGGCAG

[1763] CCGACTTCGTGCTGATTCCGGTGCAGCCAAGCCCTTACGACATATGGGCCAC

[1764] CGCCGACCTGGTGGAGCTGGTTAAGCAGCGCATTGAGGTCACGGATGGAAG

[1765] GCTACAAGCGGCCTTTGTCGTGTCGCGGGCGATCAAAGGCACGCGCATCGG

[1766] CGGTGAGGTTGCCGAGGCGCTGGCCGGGTACGAGCTGCCCATTCTTGAGTC

[1767] CCGTATCACGCAGCGCGTGAGCTACCCAGGCACTGCCGCCGCCGGCACAAC

[1768] CGTTCTTGAATCAGAACCCGAGGGCGACGCTGCCCGCGAGGTCCAGGCGCT

[1769] GGCCGCTGAAATTAAATCAAAACTCATTTGAGTTAATGAGGTAAAGAGAAAAT

[1770] GAGCAAAAGCACAAACACGCTAAGTGCCGGCCGTCCGAGCGCACGCAGCAG

[1771] CAAGGCTGCAACGTTGGCCAGCCTGGCAGACACGCCAGCCATGAAGCGGGT

[1772] CAACTTTCAGTTGCCGGCGGAGGATCACACCAAGCTGAAGATGTACGCGGTA

[1773] CGCCAAGGCAAGACCATTACCGAGCTGCTATCTGAATACATCGCGCAGCTAC

[1774] CAGAGTAAATGAGCAAATGAATAAATGAGTAGATGAATTTTAGCGGCTAAAGG

[1775] AGGCGGCATGGAAAATCAAGAACAACCAGGCACCGACGCCGTGGAATGCCC CATGTGTGGAGGAACGGGCGGTTGGCCAGGCGTAAGCGGCTGGGTTGCCT

[1776] GCCGGCCCTGCAATGGCACTGGAACCCCCAAGCCCGAGGAATCGGCGTGA

[1777] GCGGTCGCAAACCATCCGGCCCGGTACAAATCGGCGCGGCGCTGGGTGATG

[1778] ACCTGGTGGAGAAGTTGAAGGCCGCGCAGGCCGCCCAGCGGCAACGCATC

[1779] GAGGCAGAAGCACGCCCCGGTGAATCGTGGCAAGCGGCCGCTGATCGAATC

[1780] CGCAAAGAATCCCGGCAACCGCCGGCAGCCGGTGCGCCGTCGATTAGGAAG

[1781] CCGCCCAAGGGCGACGAGCAACCAGATTTTTTCGTTCCGATGCTCTATGACG

[1782] TGGGCACCCGCGATAGTCGCAGCATCATGGACGTGGCCGTTTTCCGTCTGTC

[1783] GAAGCGTGACCGACGAGCTGGCGAGGTGATCCGCTACGAGCTTCCAGACGG

[1784] GCACGTAGAGGTTTCCGCAGGGCCGGCCGGCATGGCCAGTGTGTGGGATTA

[1785] CGACCTGGTACTGATGGCGGTTTCCCATCTAACCGAATCCATGAACCGATAC

[1786] CGGGAAGGGAAGGGAGACAAGCCCGGCCGCGTGTTCCGTCCACACGTTGC

[1787] GGACGTACTCAAGTTCTGCCGGCGAGCCGATGGCGGAAAGCAGAAAGACGA

[1788] CCTGGTAGAAACCTGCATTCGGTTAAACACCACGCACGTTGCCATGCAGCGT

[1789] ACGAAGAAGGCCAAGAACGGCCGCCTGGTGACGGTATCCGAGGGTGAAGCC

[1790] TTGATTAGCCGCTACAAGATCGTAAAGAGCGAAACCGGGCGGCCGGAGTAC

[1791] ATCGAGATCGAGCTAGCTGATTGGATGTACCGCGAGATCACAGAAGGCAAGA

[1792] ACCCGGACGTGCTGACGGTTCACCCCGATTACTTTTTGATCGATCCCGGCAT

[1793] CGGCCGTTTTCTCTACCGCCTGGCACGCCGCGCCGCAGGCAAGGCAGAAGC

[1794] CAGATGGTTGTTCAAGACGATCTACGAACGCAGTGGCAGCGCCGGAGAGTT

[1795] CAAGAAGTTCTGTTTCACCGTGCGCAAGCTGATCGGGTCAAATGACCTGCCG

[1796] GAGTACGATTTGAAGGAGGAGGCGGGGCAGGCTGGCCCGATCCTAGTCATG

[1797] CGCTACCGCAACCTGATCGAGGGCGAAGCATCCGCCGGTTCCTAATGTACG

[1798] GAGCAGATGCTAGGGCAAATTGCCCTAGCAGGGGAAAAAGGTCGAAAAGGT

[1799] CTCTTTCCTGTGGATAGCACGTACATTGGGAACCCAAAGCCGTACATTGGGA

[1800] ACCGGAACCCGTACATTGGGAACCCAAAGCCGTACATTGGGAACCGGTCAC

[1801] ACATGTAAGTGACTGATATAAAAGAGAAAAAAGGCGATTTTTCCGCCTAAAAC

[1802] TCTTTAAAACTTATTAAAACTCTTAAAACCCGCCTGGCCTGTGCATAACTGTCT

[1803] GGCCAGCGCACAGCCGAAGAGCTGCAAAAAGCGCCTACCCTTCGGTCGCTG

[1804] CGCTCCCTACGCCCCGCCGCTTCGCGTCGGCCTATCGCGGCCGCTGGCCG

[1805] CTCAAAAATGGCTGGCCTACGGCCAGGCAATCTACCAGGGCGCGGACAAGC

[1806] CGCGCCGTCGCCACTCGACCGCCGGCGCCCACATCAAGGCACCCTGCCTCG

[1807] CGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCGGAGAC

[1808] GGTCACAGCTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGCCCGTCAGGG

[1809] CGCGTCAGCGGGTGTTGGCGGGTGTCGGGGCGCAGCCATGACCCAGTCAC

[1810] GTAGCGATAGCGGAGTGTATACTGGCTTAACTATGCGGCATCAGAGCAGATT

[1811] GTACTGAGAGTGCACCATATGCGGTGTGAAATACCGCACAGATGCGTAAGGA

[1812] GAAAATACCGCATCAGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCG CTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAAT

[1813] ACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAA

[1814] GGCCAGCAAA

[1815] UBQ10 promotor (Arabidopsis Thaliana);

[1816] SEQ ID NO: 32:

[1817] TCGAGCTGCAGGTCAACGGATCAGGATATTCTTGTTTAAGATGTTGAACTCTA

[1818] TGGAGGTTTGTATGAACTGATGATCTAGGACCGGATAAGTTCCCTTCTTCATA

[1819] GCGAACTTATTCAAAGAATGTTTTGTGTATCATTCTTGTTACATTGTTATTAATG

[1820] AAAAAATATTATTGGTCATTGGACTGAACACGAGTGTTAAATATGGACCAGGC

[1821] CCCAAATAAGATCCATTGATATATGAATTAAATAACAAGAATAAATCGAGTCAC

[1822] CAAACCACTTGCCTTTTTTAACGAGACTTGTTCACCAACTTGATACAAAAGTCA

[1823] TTATCCTATGCAAATCAATAATCATACAAAAATATCCAATAACACTAAAAAATTA

[1824] AAAGAAATGGATAATTTCACAATATGTTATACGATAAAGAAGTTACTTTTCCAA

[1825] GAAATTCACTGATTTTATAAGCCCACTTGCATTAGATAAATGGCAAAAAAAAAC

[1826] AAAAAGGAAAAGAAATAAAGCACGAAGAATTCTAGAAAATACGAAATACGCTT

[1827] CAATGCAGTGGGACCCACGGTTCAATTATTGCCAATTTTCAGCTCCACCGTAT

[1828] ATTTAAAAAATAAAACGATAATGCTAAAAAAATATAAATCGTAACGATCGTTAA

[1829] ATCTCAACGGCTGGATCTTATGACGACCGTTAGAAATTGTGGTTGTCGACGA

[1830] GTCAGTAATAAACGGCGTCAAAGTGGTTGCAGCCGGCACACACGAGTCGTGT

[1831] TTATCAACTCAAAGCACAAATACTTTTCCTCAACCTAAAAATAAGGCAATTAGC

[1832] CAAAAACAACTTTGCGTGTAAACAACGCTCAATACACGTGTCATTTTATTATTA

[1833] GCTATTGCTTCACCGCCTTAGCTTTCTCGTGACCTAGTCGTCCTCGTCTTTTC

[1834] TTCTTCTTCTTCTATAAAACAATACCCAAAGAGCTCTTCTTCTTCACAATTCAG

[1835] ATTTCAATTTCTCAAAATCTTAAAAACTTTCTCTCAATTCTCTCTACCGTGATCA

[1836] AGGTAAATTTCTGTGTTCCTTATTCTCTCAAAATCTTCGATTTTGTTTTCGTTCG

[1837] ATCCCAATTTCGTATATGTTCTTTGGTTTAGATTCTGTTAATCTTAGATCGAAC

[1838] ACGATTTTCTGGGTTTGATCGTTAGATATCATCTTAATTCTCGATTAGGGTTTC

[1839] ATAGATATCATCCGATTTGTTCAAATAATTTGAGTTTTGTCGAATAATTACTCTT

[1840] CGATTTGTGATTTCTATCTAGATCTGGTGTTAGTTTCTAGTTTGTGCGATCGAA

[1841] TTTGTCGATTAATCTGAGTTTTTCTGATTAACAGG

[1842] Col1A1 in pET151 ;

[1843] SEQ ID NO: 33:

[1844] TAAAAGGGCGAGCTCAGATCCGGCTGCTAACAAAGCCCGAAAGGAAGCTGA

[1845] GTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCT

[1846] AAACGGGTCTTGAGGGGTTTTTTGCTGAAAGGAGGAACTATATCCGGATATC

[1847] CCGCAAGAGGCCCGGCAGTACCGGCATAACCAAGCCTATGCCTACAGCATC CAGGGTGACGGTGCCGAGGATGACGATGAGCGCATTGTTAGATTTCATACAC

[1848] GGTGCCTGACTGCGTTAGCAATTTAACTGTGATAAACTACCGCATTAAAGCTA

[1849] GCTTATCGATGATAAGCTGTCAAACATGAGAATTAATTCTTGAAGACGAAAGG

[1850] GCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTT

[1851] AGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTT

[1852] ATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATA

[1853] AATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGT

[1854] CGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGA

[1855] AACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGT

[1856] TACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCG

[1857] AAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTA

[1858] TTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATT

[1859] CTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGAT

[1860] GGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACA

[1861] CTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCG

[1862] CTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACC

[1863] GGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGC

[1864] AGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAG

[1865] CTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACC

[1866] ACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGA

[1867] GCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGT

[1868] AAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGG

[1869] ATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTG

[1870] GTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCA

[1871] TTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAA

[1872] AATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGA

[1873] TCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAA

[1874] CAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACC

[1875] AACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTG

[1876] TCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACC

[1877] GCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGC

[1878] GATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGG

[1879] CGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAG

[1880] CGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCG

[1881] CCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGG

[1882] GTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTA

[1883] TCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGT

[1884] GATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCT TTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCG TTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATAC CGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAG CGGAAGAGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTC ACACCGCAATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAA GCCAGTATACACTCCGCTATCGCTACGTGACTGGGTCATGGCTGCGCCCCGA CACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCA TCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGT TTTCACCGTCATCACCGAAACGCGCGAGGCAGCTGCGGTAAAGCTCATCAGC GTGGTCGTGAAGCGATTCACAGATGTCTGCCTGTTCATCCGCGTCCAGCTCG TTGAGTTTCTCCAGAAGCGTTAATGTCTGGCTTCTGATAAAGCGGGCCATGTT AAGGGCGGTTTTTTCCTGTTTGGTCACTGATGCCTCCGTGTAAGGGGGATTT CTGTTCATGGGGGTAATGATACCGATGAAACGAGAGAGGATGCTCACGATAC GGGTTACTGATGATGAACATGCCCGGTTACTGGAACGTTGTGAGGGTAAACA ACTGGCGGTATGGATGCGGCGGGACCAGAGAAAAATCACTCAGGGTCAATG CCAGCGCTTCGTTAATACAGATGTAGGTGTTCCACAGGGTAGCCAGCAGCAT CCTGCGATGCAGATCCGGAACATAATGGTGCAGGGCGCTGACTTCCGCGTTT CCAGACTTTACGAAACACGGAAACCGAAGACCATTCATGTTGTTGCTCAGGT

[1885] CGCAGACGTTTTGCAGCAGCAGTCGCTTCACGTTCGCTCGCGTATCGGTGAT TCATTCTGCTAACCAGTAAGGCAACCCCGCCAGCCTAGCCGGGTCCTCAACG ACAGGAGCACGATCATGCGCACCCGTGGCCAGGACCCAACGCTGCCCGAGA TGCGCCGCGTGCGGCTGCTGGAGATGGCGGACGCGATGGATATGTTCTGCC AAGGGTTGGTTTGCGCATTCACAGTTCTCCGCAAGAATTGATTGGCTCCAATT CTTGGAGTGGTGAATCCGTTAGCGAGGTGCCGCCGGCTTCCATTCAGGTCG AGGTGGCCCGGCTCCATGCACCGCGACGCAACGCGGGGAGGCAGACAAGG TATAGGGCGGCGCCTACAATCCATGCCAACCCGTTCCATGTGCTCGCCGAG GCGGCATAAATCGCCGTGACGATCAGCGGTCCAATGATCGAAGTTAGGCTG GTAAGAGCCGCGAGCGATCCTTGAAGCTGTCCCTGATGGTCGTCATCTACCT GCCTGGACAGCATGGCCTGCAACGCGGGCATCCCGATGCCGCCGGAAGCG AGAAGAATCATAATGGGGAAGGCCATCCAGCCTCGCGTCGCGAACGCCAGC AAGACGTAGCCCAGCGCGTCGGCCGCCATGCCGGCGATAATGGCCTGCTTC TCGCCGAAACGTTTGGTGGCGGGACCAGTGACGAAGGCTTGAGCGAGGGC GTGCAAGATTCCGAATACCGCAAGCGACAGGCCGATCATCGTCGCGCTCCA GCGAAAGCGGTCCTCGCCGAAAATGACCCAGAGCGCTGCCGGCACCTGTCC TACGAGTTGCATGATAAAGAAGACAGTCATAAGTGCGGCGACGATAGTCATG

[1886] CCCCGCGCCCACCGGAAGGAGCTGACTGGGTTGAAGGCTCTCAAGGGCATC GGTCGAGATCCCGGTGCCTAATGAGTGAGCTAACTTACATTAATTGCGTTGC GCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATG AATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCCAGGGTG

[1887] GTTTTTCTTTTCACCAGTGAGACGGGCAACAGCTGATTGCCCTTCACCGCCT

[1888] GGCCCTGAGAGAGTTGCAGCAAGCGGTCCACGCTGGTTTGCCCCAGCAGGC

[1889] GAAAATCCTGTTTGATGGTGGTTAACGGCGGGATATAACATGAGCTGTCTTC

[1890] GGTATCGTCGTATCCCACTACCGAGATATCCGCACCAACGCGCAGCCCGGA

[1891] CTCGGTAATGGCGCGCATTGCGCCCAGCGCCATCTGATCGTTGGCAACCAG

[1892] CATCGCAGTGGGAACGATGCCCTCATTCAGCATTTGCATGGTTTGTTGAAAAC

[1893] CGGACATGGCACTCCAGTCGCCTTCCCGTTCCGCTATCGGCTGAATTTGATT

[1894] GCGAGTGAGATATTTATGCCAGCCAGCCAGACGCAGACGCGCCGAGACAGA

[1895] ACTTAATGGGCCCGCTAACAGCGCGATTTGCTGGTGACCCAATGCGACCAGA

[1896] TGCTCCACGCCCAGTCGCGTACCGTCTTCATGGGAGAAAATAATACTGTTGA

[1897] TGGGTGTCTGGTCAGAGACATCAAGAAATAACGCCGGAACATTAGTGCAGGC

[1898] AGCTTCCACAGCAATGGCATCCTGGTCATCCAGCGGATAGTTAATGATCAGC

[1899] CCACTGACGCGTTGCGCGAGAAGATTGTGCACCGCCGCTTTACAGGCTTCGA

[1900] CGCCGCTTCGTTCTACCATCGACACCACCACGCTGGCACCCAGTTGATCGGC

[1901] GCGAGATTTAATCGCCGCGACAATTTGCGACGGCGCGTGCAGGGCCAGACT

[1902] GGAGGTGGCAACGCCAATCAGCAACGACTGTTTGCCCGCCAGTTGTTGTGC

[1903] CACGCGGTTGGGAATGTAATTCAGCTCCGCCATCGCCGCTTCCACTTTTTCC

[1904] CGCGTTTTCGCAGAAACGTGGCTGGCCTGGTTCACCACGCGGGAAACGGTC

[1905] TGATAAGAGACACCGGCATACTCTGCGACATCGTATAACGTTACTGGTTTCAC

[1906] ATTCACCACCCTGAATTGACTCTCTTCCGGGCGCTATCATGCCATACCGCGA

[1907] AAGGTTTTGCGCCATTCGATGGTGTCCGGGATCTCGACGCTCTCCCTTATGC

[1908] GACTCCTGCATTAGGAAGCAGCCCAGTAGTAGGTTGAGGCCGTTGAGCACC

[1909] GCCGCCGCAAGGAATGGTGCATGCAAGGAGATGGCGCCCAACAGTCCCCCG

[1910] GCCACGGGGCCTGCCACCATACCCACGCCGAAACAAGCGCTCATGAGCCCG

[1911] AAGTGGCGAGCCCGATCTTCCCCATCGGTGATGTCGGCGATATAGGCGCCA

[1912] GCAACCGCACCTGTGGCGCCGGTGATGCCGGCCACGATGCGTCCGGCGTA

[1913] GAGGATCGAGATCTCGATCCCGCGAAATTAATACGACTCACTATAGGGGAAT

[1914] TGTGAGCGGATAACAATTCCCCTCTAGAAATAATTTTGTTTAACTTTAAGAAGG

[1915] AGATATACATATGCATCATCACCATCACCATGGTAAGCCTATCCCTAACCCTC

[1916] TCCTCGGTCTCGATTCTACGGAAAACCTGTATTTTCAGGGACAATTGAGTTAT

[1917] GGATACGACGAAAAATCAACTGGAATATCAGTGCCAGGACCAATGGGTCCAT

[1918] CAGGACCTAGGGGTCTTCCAGGGCCACCTGGCGCTCCTGGTCCTCAGGGAT

[1919] TTCAAGGTCCTCCAGGTGAACCTGGAGAGCCTGGTGCTTCTGGTCCAATGGG

[1920] ACCAAGAGGCCCTCCTGGTCCACCTGGTAAAAATGGTGATGATGGTGAGGCT

[1921] GGAAAGCCTGGTCGTCCTGGAGAAAGAGGACCACCAGGACCACAGGGAGCT

[1922] AGAGGTTTGCCAGGTACAGCCGGTCTTCCAGGAATGAAAGGACACAGGGGT

[1923] TTTTCTGGTCTTGATGGTGCTAAGGGAGACGCAGGACCTGCTGGACCAAAGG GTGAACCTGGATCTCCTGGTGAAAATGGTGCTCCTGGACAAATGGGACCTAG

[1924] AGGATTGCCTGGCGAAAGAGGAAGACCTGGTGCTCCTGGACCAGCTGGAGC

[1925] TAGAGGAAATGATGGAGCTACTGGTGCTGCCGGACCACCAGGACCAACTGG

[1926] ACCAGCTGGACCACCTGGTTTTCCAGGAGCAGTTGGGGCTAAGGGAGAAGG

[1927] TGGACCACAAGGACCTAGAGGATCAGAAGGACCTCAAGGAGTTAGGGGAGA

[1928] ACCAGGTCCTCCTGGTCCTGCTGGAGCTGCTGGACCCGCTGGTAACCCAGG

[1929] AGCAGATGGACAACCTGGAGCAAAAGGGGCTAATGGTGCTCCTGGTATTGCA

[1930] GGTGCTCCTGGATTCCCTGGTGCTAGAGGACCTTCAGGACCTCAGGGACCAT

[1931] CCGGTCCTCCAGGACCAAAGGGAAACTCTGGTGAACCAGGGGCTCCTGGAT

[1932] CTAAGGGAGATACTGGAGCTAAGGGTGAGCCAGGACCTACAGGAATTCAAG

[1933] GACCACCAGGACCTGCAGGAGAGGAAGGTAAGAGGGGTGCTAGAGGTGAAC

[1934] CTGGACCAGCTGGATTGCCAGGCCCACCTGGAGAGAGGGGTGGTCCAGGTT

[1935] CTAGAGGTTTTCCAGGAGCTGATGGAGTTGCTGGTCCAAAGGGTCCTGCTGG

[1936] TGAGAGAGGTGCACCTGGACCAGCAGGACCTAAAGGATCACCTGGAGAAGC

[1937] TGGTCGTCCTGGTGAGGCTGGTCTTCCAGGTGCTAAGGGCCTTACAGGATCT

[1938] CCAGGATCACCAGGTCCAGATGGAAAGACTGGACCACCAGGTCCTGCAGGA

[1939] CAAGATGGAAGGCCTGGTCCACCTGGACCACCTGGTGCTAGGGGACAAGCT

[1940] GGTGTTATGGGATTCCCAGGTCCAAAGGGTGCAGCTGGTGAGCCTGGTAAA

[1941] GCTGGTGAAAGAGGAGTTCCAGGACCTCCTGGTGCTGTTGGACCTGCTGGT

[1942] AAAGATGGTGAAGCAGGTGCTCAGGGTCCACCAGGTCCAGCCGGTCCAGCT

[1943] GGTGAGAGGGGAGAGCAAGGTCCTGCTGGATCTCCTGGATTTCAAGGTTTG

[1944] CCTGGACCTGCTGGACCTCCAGGAGAGGCTGGTAAACCTGGTGAGCAGGGT

[1945] GTGCCTGGTGATCTTGGTGCTCCTGGACCTAGTGGCGCAAGAGGCGAAAGG

[1946] GGATTTCCAGGTGAAAGAGGAGTTCAGGGACCACCAGGTCCAGCTGGTCCA

[1947] AGAGGCGCTAATGGAGCTCCAGGAAATGATGGAGCAAAGGGTGATGCTGGC

[1948] GCTCCAGGAGCTCCTGGTTCTCAAGGTGCACCTGGATTGCAAGGAATGCCAG

[1949] GAGAGAGAGGTGCAGCTGGTTTGCCGGGACCAAAGGGTGATAGAGGTGATG

[1950] CTGGTCCTAAGGGAGCTGATGGAGCTCCAGGAAAGGATGGTGTGAGAGGAT

[1951] TGACTGGTCCAATTGGACCACCAGGTCCTGCTGGAGCACCAGGAGATAAAG

[1952] GAGAAGCTGGTCCTTCAGGACCAGCCGGTCCAACTGGAGCTAGAGGAGCTC

[1953] CAGGAGATAGAGGTGAACCAGGACCACCAGGACCTGCTGGGTTTGCAGGTC

[1954] CACCTGGTGCTGATGGACAACCTGGTGCTAAAGGAGAACCAGGCGATGCTG

[1955] GTGCTAAGGGAGATGCTGGCCCACCTGGTCCAGCTGGTCCTGCTGGGCCAC

[1956] CAGGTCCTATCGGAAATGTTGGTGCTCCAGGGCCTAAAGGAGCTAGAGGATC

[1957] AGCAGGTCCCCCAGGTGCTACAGGTTTTCCAGGGGCTGCTGGAAGAGTGGG

[1958] TCCCCCAGGTCCTTCTGGGAATGCAGGACCACCAGGTCCTCCAGGACCAGC

[1959] TGGAAAAGAAGGAAGTAAGGGACCAAGGGGAGAAACTGGACCTGCAGGACG

[1960] TCCAGGTGAAGTTGGTCCACCTGGACCTCCAGGTCCAGCTGGAGAGAAAGG AGCTCCAGGAGCTGATGGTCCAGCAGGTGCTCCTGGTACTCCAGGTCCACA

[1961] AGGTATTGCTGGTCAAAGGGGAGTGGTTGGTCTCCCAGGTCAGAGAGGTGA

[1962] AAGAGGTTTCCCAGGTTTGCCTGGACCATCTGGAGAACCAGGTAAGCAGGG

[1963] CCCATCTGGAGCTTCTGGTGAGAGAGGGCCACCTGGACCTATGGGACCTCC

[1964] TGGGCTTGCTGGACCTCCAGGGGAATCTGGTAGGGAGGGTGCTCCAGGTGC

[1965] TGAAGGATCACCTGGTAGAGATGGATCACCTGGAGCTAAAGGTGATAGAGG

[1966] GGAAACAGGTCCAGCAGGTCCACCTGGCGCTCCTGGTGCTCCAGGAGCACC

[1967] AGGGCCAGTTGGACCTGCTGGAAAGTCTGGAGATAGGGGAGAGACTGGACC

[1968] AGCTGGACCTGCTGGACCAATCGGTCCTGTTGGCGCTAGAGGTCCTGCTGG

[1969] ACCACAAGGTCCTAGGGGAGATAAAGGTGAAACAGGTGAACAAGGAGATAG

[1970] AGGAATTAAAGGTCATAGAGGGTTTTCTGGTTTGCAAGGACCTCCAGGACCT

[1971] CCTGGATCTCCAGGAGAGCAAGGACCATCTGGTGCAAGCGGACCTGCTGGT

[1972] CCTAGAGGACCTCCAGGTTCAGCTGGCTCACCTGGTAAGGATGGATTAAATG

[1973] GTTTGCCTGGACCAATAGGTCCACCAGGACCTAGAGGAAGGACAGGTGATG

[1974] CAGGACCAGCTGGTCCACCTGGTCCACCAGGACCACCAGGACCACCAGGTC

[1975] CACCTTCTGGTGGATATGATCTTAGTTTTCTTCCTCAACCTCCCCAGGAGAAA

[1976] GCTCATGATGGTGGTAGATATTATAGGGCT

[1977] Bovine Col1A1 expression vector VB;

[1978] SEQ ID NO: 34:

[1979] CAACTTTGTATAGAAAAGTTGAGATCTAACATCCAAAGACGAAAGGTTGAATG

[1980] AAACCTTTTTGCCATCCGACATCCACAGGTCCATTCTCACACATAAGTGCCAA

[1981] ACGCAACAGGAGGGGATACACTAGCAGCAGACCGTTGCAAACGCAGGACCT

[1982] CCACTCCTCTTCTCCTCAACACCCACTTTTGCCATCGAAAAACCAGCCCAGTT

[1983] ATTGGGCTTGATTGGAGCTCGCTCATTCCAATTCCTTCTATTAGGCTACTAAC

[1984] ACCATGACTTTATTAGCCTGTCTATCCTGGCCCCCCTGGCGAGGTTCATGTTT

[1985] GTTTATTTCCGAATGCAACAAGCTCCGCATTACACCCGAACATCACTCCAGAT

[1986] GAGGGCTTTCTGAGTGTGGGGTCAAATAGTTTCATGTTCCCCAAATGGCCCA

[1987] AAACTGACAGTTTAAACGCTGTCTTGGAACCTAATATGACAAAAGCGTGATCT

[1988] CATCCAAGATGAACTAAGTTTGGTTCGTTGAAATGCTAACGGCCAGTTGGTCA

[1989] AAAAGAAACTTCCAAAAGTCGGCATACCGTTTGTCTTGTTTGGTATTGATTGA

[1990] CGAATGCTCAAAAATAATCTCATTAATGCTTAGCGCAGTCTCTCTATCGCTTCT

[1991] GAACCCCGGTGCACCTGTGCCGAAACGCAAATGGGGAAACACCCGCTTTTT

[1992] GGATGATTATGCATTGTCTCCACATTGTATGCTTCCAAGATTCTGGTGGGAAT

[1993] ACTGCTGATAGCCTAACGTTCATGATCAAAATTTAACTGTTCTAACCCCTACTT

[1994] GACAGCAATATATAAACAGAAGGAAGCTGCCCTGTCTTAAACCTTTTTTTTTAT

[1995] CATCATTATTAGCTTACTTTCATAATTGCGACTGGTTCCAATTGACAAGCTTTT

[1996] GATTTTAACGACTTTTAACGACAACTTGAGAAGATCAAAAAACAACTAATTATT CGAAACGACAAGTTTGTACAAAAAAGCAGGCTGCCACCATGAGATTTCCTTCA

[1997] ATTTTTACTGCTGTTTTATTCGCAGCATCCTCCGCATTAGCTGCTCCAGTCAA

[1998] CACTACAACAGAAGATGAAACGGCACAAATTCCGGCTGAAGCTGTCATCGGT

[1999] TACTCAGATTTAGAAGGGGATTTCGATGTTGCTGTTTTGCCATTTTCCAACAG

[2000] CACAAATAACGGGTTATTGTTTATAAATACTACTATTGCCAGCATTGCTGCTAA

[2001] AGAAGAAGGGGTATCTCTCGAGAAAAGAGAGGCTGAAGCTCATCACCATCAC

[2002] CATCACGAAAACCTGTATTTTCAGGGCCAATTGTCTTATGGTTACGATGAAAA

[2003] GTCCACTGGTATTTCTGTCCCTGGACCAATGGGTCCATCAGGTCCTAGAGGT

[2004] TTGCCAGGTCCTCCAGGTGCTCCAGGTCCACAAGGTTTTCAAGGTCCACCAG

[2005] GTGAACCAGGAGAACCCGGTGCTTCTGGTCCAATGGGTCCAAGAGGTCCAC

[2006] CTGGACCTCCTGGTAAGAACGGTGATGATGGTGAGGCTGGTAAGCCAGGAA

[2007] GACCTGGAGAAAGAGGACCACCTGGTCCACAAGGAGCTAGAGGTTTACCCG

[2008] GTACAGCTGGTTTGCCTGGTATGAAGGGTCATAGAGGTTTCTCCGGTTTGGA

[2009] TGGAGCTAAGGGTGATGCTGGACCTGCTGGTCCTAAGGGAGAGCCAGGATC

[2010] TCCAGGTGAGAACGGAGCTCCTGGACAAATGGGACCAAGAGGTCTGCCTGG

[2011] TGAAAGAGGTAGACCAGGTGCTCCTGGTCCTGCCGGAGCCAGAGGTAATGA

[2012] TGGTGCTACTGGTGCTGCTGGTCCTCCAGGTCCAACTGGTCCTGCTGGTCCT

[2013] CCAGGTTTTCCAGGTGCTGTTGGAGCCAAAGGTGAAGGTGGCCCACAAGGT

[2014] CCTAGAGGATCTGAAGGTCCACAGGGTGTTAGAGGTGAACCAGGTCCACCA

[2015] GGTCCAGCTGGAGCTGCTGGTCCTGCTGGTAACCCAGGTGCAGATGGTCAA

[2016] CCAGGTGCTAAGGGTGCCAACGGTGCTCCAGGCATTGCTGGAGCTCCAGGT

[2017] TTTCCAGGAGCTAGAGGTCCTTCTGGACCTCAAGGACCATCTGGTCCACCTG

[2018] GACCAAAAGGTAACTCTGGAGAACCAGGTGCTCCTGGTTCCAAAGGTGATAC

[2019] CGGTGCTAAAGGAGAGCCAGGTCCTACTGGTATTCAAGGTCCTCCTGGTCCT

[2020] GCTGGTGAAGAGGGTAAAAGAGGAGCCAGAGGTGAACCTGGTCCAGCCGGT

[2021] TTGCCAGGTCCACCAGGTGAAAGAGGAGGTCCAGGATCTAGAGGTTTTCCAG

[2022] GTGCTGACGGTGTTGCTGGTCCAAAGGGTCCTGCTGGTGAGAGAGGAGCCC

[2023] CAGGTCCAGCTGGACCTAAAGGATCTCCAGGAGAAGCTGGTCGTCCTGGTG

[2024] AAGCTGGTCTTCCAGGTGCTAAGGGTTTGACAGGCTCCCCAGGATCCCCTGG

[2025] TCCAGATGGTAAGACTGGACCACCTGGACCAGCTGGTCAAGATGGAAGACC

[2026] AGGTCCTCCAGGACCACCTGGAGCCAGAGGCCAAGCTGGTGTTATGGGTTT

[2027] CCCTGGTCCAAAAGGTGCTGCTGGAGAACCGGGTAAAGCCGGTGAAAGAGG

[2028] TGTTCCAGGTCCTCCAGGTGCTGTTGGTCCTGCTGGTAAAGATGGAGAAGCC

[2029] GGTGCTCAAGGACCACCAGGTCCAGCTGGTCCAGCTGGAGAGAGAGGTGAA

[2030] CAAGGTCCTGCTGGATCCCCAGGTTTCCAAGGTTTGCCTGGTCCTGCTGGAC

[2031] CACCAGGTGAAGCTGGTAAGCCAGGTGAACAAGGAGTTCCAGGTGATTTGG

[2032] GTGCTCCTGGTCCATCCGGTGCTAGAGGTGAAAGAGGATTTCCAGGAGAGA

[2033] GAGGTGTTCAAGGACCACCTGGTCCTGCTGGACCAAGAGGAGCAAACGGTG CTCCAGGAAACGATGGTGCTAAAGGTGATGCTGGTGCTCCAGGAGCTCCAG

[2034] GTTCACAAGGTGCTCCAGGTTTGCAAGGAATGCCTGGAGAAAGAGGTGCTG

[2035] CTGGATTACCAGGTCCAAAAGGAGACAGAGGTGATGCCGGTCCAAAGGGTG

[2036] CTGATGGTGCTCCAGGTAAGGACGGTGTGCGAGGTTTGACTGGACCAATTG

[2037] GACCTCCAGGTCCAGCTGGAGCCCCAGGAGATAAGGGTGAAGCTGGTCCAT

[2038] CGGGTCCTGCTGGTCCTACTGGTGCTAGAGGAGCTCCAGGAGATAGAGGTG

[2039] AACCTGGTCCACCTGGACCCGCTGGATTTGCTGGTCCACCAGGTGCTGATG

[2040] GTCAACCAGGGGCTAAAGGTGAACCAGGTGACGCTGGAGCTAAAGGAGATG

[2041] CCGGTCCACCAGGTCCAGCCGGTCCAGCTGGTCCTCCTGGTCCAATTGGTA

[2042] ATGTTGGCGCTCCTGGTCCTAAGGGTGCCAGAGGTTCAGCTGGACCACCAG

[2043] GTGCTACTGGTTTCCCAGGTGCTGCCGGTAGAGTTGGTCCACCAGGTCCATC

[2044] TGGTAACGCTGGTCCTCCAGGTCCACCAGGTCCTGCTGGAAAGGAGGGATC

[2045] TAAGGGACCAAGAGGTGAAACTGGTCCAGCTGGAAGACCTGGTGAAGTTGG

[2046] TCCTCCAGGACCACCAGGTCCTGCTGGAGAAAAAGGTGCTCCAGGTGCTGAT

[2047] GGACCAGCTGGTGCTCCTGGTACCCCAGGTCCACAAGGTATTGCTGGTCAAA

[2048] GAGGTGTGGTTGGTCTTCCTGGACAAAGAGGTGAGAGAGGTTTTCCTGGTTT

[2049] GCCAGGTCCTTCTGGAGAACCTGGAAAACAGGGTCCCTCTGGTGCTTCTGGT

[2050] GAGAGAGGTCCTCCAGGACCAATGGGACCTCCAGGTTTGGCTGGTCCACCA

[2051] GGTGAGTCTGGTAGAGAGGGTGCTCCAGGTGCTGAGGGTTCTCCAGGTAGA

[2052] GATGGTAGTCCAGGTGCTAAAGGAGACAGAGGAGAGACTGGTCCAGCTGGT

[2053] CCTCCAGGTGCTCCTGGTGCTCCCGGTGCTCCTGGTCCTGTCGGTCCTGCT

[2054] GGAAAGTCTGGTGATAGAGGTGAGACTGGTCCAGCTGGACCAGCTGGTCCA

[2055] ATCGGACCAGTTGGAGCTAGAGGTCCTGCTGGTCCTCAAGGTCCAAGAGGT

[2056] GACAAGGGTGAAACAGGTGAGCAAGGTGACAGAGGTATTAAAGGACATAGA

[2057] GGTTTCTCTGGTCTACAGGGACCTCCTGGTCCTCCAGGTTCCCCAGGTGAGC

[2058] AAGGTCCTTCTGGTGCTTCAGGTCCTGCTGGTCCAAGAGGTCCACCAGGTTC

[2059] TGCTGGTTCTCCAGGAAAGGATGGTTTGAATGGTTTGCCTGGACCTATTGGT

[2060] CCTCCTGGACCTAGAGGTAGAACAGGTGATGCTGGTCCTGCTGGTCCACCTG

[2061] GTCCTCCTGGTCCTCCTGGTCCCCCTGGACCTCCATCTGGAGGTTACGATTT

[2062] GTCTTTTTTGCCACAACCTCCACAGGAAAAAGCCCATGACGGTGGAAGATATT

[2063] ATAGAGCTTAAACCCAGCTTTCTTGTACAAAGTGGTGTTTGTAGCCTTAGACA

[2064] TGACTGTTCCTCAGTTCAAGTTGGGCACTTACGAGAAGACCGGTCTTGCTAG

[2065] ATTCTAATCAAGAGGATGTCAGAATGCCATTTGCCTGAGAGATGCAGGCTTCA

[2066] TTTTTGATACTTTTTTATTTGTAACCTATATAGTATAGGATTTTTTTTGTCATTTT

[2067] GTTTCTTCTCGTACGAGCTTGCTCCTGATCAGCCTATCTCGCAGCTGATGAAT

[2068] ATCTTGTGGTAGGGGTTTGGGAAAATCATTCGAGTTTGATGTTTTTCTTGGTA

[2069] TTTCCCACTCCTCTTCAGAGTACAGAAGATTAAGTGAGACCTTCGTTTGTGCG

[2070] GATCCCCCACACACCATAGCTTCAAAATGTTTCTACTCCTTTTTTACTCTTCCA GATTTTCTCGGACTCCGCGCATCGCCGTACCACTTCAAAACACCCAAGCACA GCATACTAAATTCCCCCTCTTTCTTCCTCTAGGGTGTCGTTAATTACCCGTACT AAAGGTTTGGAAAAGAAAAAAGAGACCGCCTCGTTTCTTTTTCTTCGTCGAAA AAGGCAATAAAAATTTTTATCACGTTTCTTTTTCTTGAAAATTTTTTTTTTTGATT TTTTTCTCTTTCGATGACCTCCCATTGATATTTAAGTTAATAAATGGTCTTCAAT TTCTCAAGTTTCAGTTTCATTTTTCTTGTTCTATTACAACTTTTTTTACTTCTTGC TCATTAGAAAGAAAGCATAGCAATCTAATCTAAGGGCGGTGTTGACAATTAAT CATCGGCATAGTATATCGGCATAGTATAATACGACAAGGTGAGGAACTAAAC CATGGCCAAGTTGACCAGTGCCGTTCCGGTGCTCACCGCGCGCGACGTCGC CGGAGCGGTCGAGTTCTGGACCGACCGGCTCGGGTTCTCCCGGGACTTCGT GGAGGACGACTTCGCCGGTGTGGTCCGGGACGACGTGACCCTGTTCATCAG CGCGGTCCAGGACCAGGTGGTGCCGGACAACACCCTGGCCTGGGTGTGGG TGCGCGGCCTGGACGAGCTGTACGCCGAGTGGTCGGAGGTCGTGTCCACGA ACTTCCGGGACGCCTCCGGGCCGGCCATGACCGAGATCGGCGAGCAGCCG TGGGGGCGGGAGTTCGCCCTGCGCGACCCGGCCGGCAACTGCGTGCACTT CGTGGCCGAGGAGCAGGACTGACACGTCCGACGCGGCCCGACGGGTCCGA GGCCTCGGAGATCCGTCCCCCTTTTCCTTTGTCGATATCATGTAATTAGTTAT GTCACGCTTACATTCACGCCCTCCCCCCACATCCGCTCTAACCGAAAAGGAA GGAGTTAGACAACCTGAAGTCTAGGTCCCTATTTATTTTTTTATAGTTATGTTA GTATTAAGAACGTTATTTATATTTCAAATTTTTCTTTTTTTTCTGTACAGACGCG TGTACGCATGTAACATTATACTGAAAACCTTGCTTGAGAAGGTTTTGGGACGC TCGAAGGCTTTAATTTGCAAGCTGGAGACCAACATGTGAGCAAAAGGCCAGC AAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCT CCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCG AAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTC GTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTC TCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAG TTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTT CAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGG TAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAG AGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTAC GGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTA CCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGG TAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGAT CTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAA AACTCACGTTAAGGGATTTTGGTCATGAGATC Table 1. Further sequences that may be used in the context of the present invention as collagen as expressed by a non-anima organism

[2071]

[2072]

[2073]

[2074]

[2075]

[2076]

[2077]

[2078]

[2079]

[2080]

[2081]

[2082]

[2083]

[2084]

[2085]

[2086]

[2087]

[2088]

[2089]

[2090]

[2091]

[2092]

[2093]

[2094]

[2095]

[2096]

[2097]

[2098]

[2099]

[2100]

[2101]

[2102]

[2103]

[2104]

[2105]

[2106]

[2107]

[2108]

[2109]

[2110]

[2111]

[2112]

[2113]

[2114]

[2115]

[2116] Table 2. Strains that may be used in the context of the present invention as non-animal organism (DSP Host organisms and example of expressable collagens. This list only mentions strains that were generated for recombinant protein production. E. coli JM109 that i also usable to amplify and purify plasmids for further applications is not enlisted):

[2117] In a preferred embodiment, the collagen expressed by the non-animal organism is selected from Table 1 .

[2118] In a preferred embodiment, the non-animal organism is selected from those depicted in Table 2 - which may be independent of the exemplified collagen.

[2119] The non-animal organism preferably contains genetic material that encodes for the collagen that naturally occurs extracellularly in one or more animal species. Such genetic material is a foreign genetic material. Its sequence at least partly corresponds to a sequence of animal origin, while it is expressed in a non-animal organism. The genetic material may be any genetic material known in the art. The genetic material may, for instance, be deoxyribonucleic acid (DNA), ribonucleic acid (RNA), an analogue thereof such as peptide nucleic acid (PNA), morpholino and locked nucleic acid (LNA), glycol nucleic acid (GNA), threose nucleic acid (TNA) and hexitol nucleic acids (HNA), or a combination of two or more thereof. It will be understood that the DNA, RNA, analogue thereof, or combination thereof may be optionally stabilized by any means such as, e.g., terminal (e.g., 3’-terminal and / or 5’ terminal) capping.

[2120] DNA may be double stranded DNA or single stranded DNA, preferably double stranded DNA. RNA may be double stranded RNA or single stranded RNA. Also, a nucleic acid analogue may be double stranded or single stranded. DNA, RNA, analogue thereof, or combination thereof may be linear or circular. For instance, circular DNA may be a plasmid.

[2121] The present invention is not limited to a certain type of gene vector. The present invention is not limited to a certain type of plasmid.

[2122] A large variety of examples of plasmids that may be used in the context of the present invention are well-known by the person skilled in the art. It will be understood that the plasmid, including its enhancers and optional enhancer sequences, start and stop sequences may be adapted to the non-animal organism as used. Optionally, a plasmid or other vector may further lead to resistance against one or more antibiotics. This allows selection of successfully transfected cells from nontransfected cells. While non-transfected cells are killed or at least growth-arrested, the transfected cells are viable in a culture medium containing the respective antibiotic. This is well-known by the person skilled in the art. Examples that may be used in the context of the present invention may be selected from Tables 1 and 2 herein, without being bound to certain collagen cargos, which can be freely adapted by routine means.

[2123] The inclusion of DNA, RNA, analogue thereof, or combination thereof into the nonanimal organism may also be considered as “transfection”. Such transfection may be achieved by any means known in the art such as, e.g., electroporation, using a gene gun or a vector (e.g., a viral vector, a cell-penetrating peptide, etc.). A transfection may be a permanent or a transient transfection. For the expression in vectors with N-terminal tags, such as, e.g., pET151 , a start codon may be omitted, because there may be already one before the tag. Methionin as the first amino acid may optionally be cleaved anyway. However, when it is desirable, the start codon may be kept, because there is only one RBS, before the tag, so there should be no recognition of two different open reading frames (ORFs). However, pET151 may need one or two stop codons. Optionally, two TAAs may be added at the end of the sequence.

[2124] When the non-animal organism is a plant, transfection may also involve the generation of a callus. Optionally, the cell wall of a cell to be transfected may be partly or completely removed.

[2125] The person skilled in the art is well aware of a variety of methods for heterologous expression of genes in non-animal organisms as such. Examples are mentioned below in the example section.

[2126] In a transient transfection, the genetic material is incorporated into the organism for a limited time such as for few hours, few days, few weeks or few months. A permanent transfection leads to a longer duration of the genetic material in the cells. Typically, a non-animal organism that expresses the genetic material encoding for the collagen permanently can also pass the genetic material to subsequent generations upon breeding. Optionally, DNA, RNA, analogue thereof, or combination thereof, in particular DNA, may be incorporated into the non-animal organism’s genome. This may be preferably passed to subsequent generations of the non-animal organism. A permanent expression may also be achieved by using one or more plasmids that may optionally replicate in the cells of the non-animal organism expressing the collagen. In a preferred embodiment, the non-animal organism comprises the genetic material encoding for the collagen permanently. In a preferred embodiment, the genetic material is incorporated in the non-animal organism’s genome. In a preferred embodiment, the non-animal organism comprises the genetic material encoding for the collagen permanently and the genetic material is incorporated in the non-animal organism’s genome.

[2127] In an alternative preferred embodiment, the non-animal organism comprises the genetic material encoding for the collagen transiently.

[2128] In a preferred embodiment, the non-animal organism as a whole is comestible, i.e. , edible and / or drinkable and / or suitable to prepare a beverage or foodstuff of it.

[2129] As used in the context of the present invention, the term “comestible” may be understood as generally understood in the art as consumable without (substantial) health risk when consuming reasonable amounts as indicated herein. Preferably, “comestible” means components suitable for animal, and in particular human, food consumption. This may also be interpreted in the context of accreditation of official regulatory offices. As used in this context, “reasonable amounts” may depend on the properties of the material. Typically, a reasonable consumable amount may range in the oral dose range of the collagen of milligrams or grams per day, such as, e.g., 0.0001 to 1000 g / day of the collagen, 0.005 to 100 g / day of the collagen, 0.001 to 10 g / day of the collagen, 0.05 to 5 g / day of the collagen, or 0.1 to 1 g / day of the collagen. Specific ranges will be understood by a person skilled in the art.

[2130] In a preferred embodiment, the collagen occurs in or is used in natural animal- derived food or comestible nutrient. Preferably, such animal-based comestible nutrient is vegetarian.

[2131] In a preferred embodiment, the collagen is used as a gelling agent in food, a beverage, a medication, a capsule (e.g., a vitamin, nutrient or medicament capsule), a photographic film, paper, and / or a cosmetic (e.g., as texture conditioner, moisturizer, and / or filler).

[2132] In a preferred embodiment, the collagen occurs in or is used in a composition that is comprised in at least one vegetarian animal-based comestible nutrient, in particular a dairy and / or an egg-based product. In a preferred embodiment, the collagen occurs in or is used in a dessert such as, e.g., candy (e.g., gummy candy, marshmallows), ice cream, a bakery good (e.g., torte, sweetie, cake, baklava, cake glaze, frostings, cake fillings, no-bake cakes), a dip, a jam, a diary product (e.g., yogurt or creams, desserts). In a preferred embodiment, the collagen occurs in or is used in a spicy food such as in aspic, a (optionally vergetarian) sausage, cream cheese, spread, margarine and / or a sauce or soup.

[2133] In a preferred embodiment, the collagen occurs in or is used in a composition that is comprised in at least one vegetarian animal-based comestible nutrient, in particular a vegetarian animal-based comestible nutrient selected from the group consisting of candy (e.g., gummy candy, marshmallows), ice cream, a bakery good (e.g., torte, sweetie, cake, baklava, cake glaze, frostings, cake fillings, no-bake cakes), a dip, a diary product (e.g., yogurt or creams, desserts ), aspic, a vegetarian sausage, cream cheese, jam, spread, margarine and / or a sauce or soup.

[2134] In a preferred embodiment, the collagen is used as a stabilizer, thickener, or texturizer such as, e.g., in food, medical uses, cosmetic uses. Collagen may also be used as binder and / or as glue.

[2135] In a preferred embodiment, the collagen is usable for cooking such as, e.g., as an instant ingredient, and may be provided as powder, granule, or sheet.

[2136] Accordingly, a “comestible nutrient” may be understood in the broadest sense as any material that is consumable without (substantial) health risk when consuming reasonable amounts. Preferably, a “comestible nutrient” may be understood in the broadest sense as any foodstuff composition that may be consumed by drinking and / or eating.

[2137] The collagen that naturally occurs extracellularly in one or more animal species may be located in the non-animal organism expressing it at any location.

[2138] The intracellular protein may be a soluble collagen or may form collagen aggregates, which may optionally precipitate.

[2139] In a preferred embodiment, the expressed collagen is located intracellularly in the non-animal organism. For instance, the expressed collagen may be located in the cytoplasm of cells of non-animal organism and / or may be located attached to a membrane of cells of nonanimal organism or included in a membrane of cells of non-animal organism. The intracellular collagen may be a soluble collagen or may be a membrane-bound collagen and / or may form collagen aggregates. Alternatively, the expressed collagen may be located in protein storage vacuoles, endoplasmic reticulum (ER) or chloroplasts.

[2140] In another preferred embodiment, the expressed collagen is located extracellularly in the non-animal organism.

[2141] The extracellular collagen may be secreted by cells of the non-animal organism. In the case that the non-animal organism is a monocellular organism such as, e.g., a bacterium or a yeast, the extracellular collagen may be secreted into the cell culture medium in which the cells are cultivated. In the case that the non-animal organism is a multicellular organism such as, e.g., a plant or a fungus, the extracellular collagen may be secreted outside the organism, e.g., via a gland, or may be located in the extracellular space inside the non-animal organism. The extracellular collagen may be a soluble collagen or may be a membrane-bound collagen and / or may form collagen aggregates.

[2142] The expressed collagen may optionally comprise one or more means for its purification.

[2143] In a preferred embodiment, the expressed collagen contains a tag, in particular a His-tag.

[2144] Such a tag may allow affinity purification of the expressed collagen. A tag may be any sequence that allows specific binding. For instance, a tag may be a His-tag, a (poly)peptide sequence that may specifically bind to a target structure such as, e.g, streptavidin, or may be a sequence that is detected by an antibody. A His-tag may comprise any number of histidine (His) residues such as, e.g., five, six, seven or eight His residues. In a preferred embodiment, the collagen comprises a hexahistidine (6His) tag that may contain six consecutive histidine residues. Such tag (e.g., a His-tag) may be located N-terminally or C-terminally of the collagen strand. A cleavage site for polypeptide purification may be any method known in the art for purifying polypeptides. For instance, a cleavage site for polypeptide purification purposes may be TEV cleavage site (as recognized by TEV (Tobacco Etch Virus) cysteine protease) or a functional homologue thereof, in particular TEV.

[2145] The collagen and optionally further parts of the non-animal organism is expressed by forming part of a comestible nutrient composition.

[2146] The obtained comestible nutrient product (e.g., a powder or a liquid or a gel) may optionally be hermetically sealed. The product may be prepared at and / or subsequently treated below room temperature (i.e., <20°C), at room temperature (e.g., (approximately) 20°C), or at increased temperature (>20°C, such as exemplarily at 60-120°C for faster dissolving). The product may optionally be sterilized (e.g., by pasteurization, heating, irradiation with ultraviolet (UV) light or X- rays, gamma-rays, etc.).

[2147] The collagen, or non-animal organism comprising it or comestible nutrient composition comprising the collagen and / or non-animal organism or parts thereof may be consumed directly. Optionally, the collagen or comestible nutrient composition comprising it may be processed further by any means known in the art. In one embodiment, the method further comprises a step of:

[2148] (a) fermenting the comestible nutrient composition;

[2149] (b) smoking the comestible nutrient composition;

[2150] (c) pickling a comestible good with a composition comprising the comestible nutrient composition, preferably in combination with one or more further component such as at least 1 % by weight, referred to the composition, of sodium chloride, at least 1 % by weight, referred to the composition, of one or more types of sugar, at least 1 % by weight, referred to the composition, of acetic acid, at least 0.5% by weight, referred to the composition, of ethanol, or at least 5% by weight, referred to the composition, of one or more types of edible oil;

[2151] (d) thickening the comestible nutrient composition to obtain a syrup, in particular wherein sugar and / or other sweetener is added to obtain a sweet syrup; and / or

[2152] (e) combining one or more collagens or collagen fragments in a comestible nutrient composition As used herein, fermenting may be understood in the broadest sense as any kind of metabolic process that produces chemical changes in the organic substrates as contained in the composition through the action of enzymes. Such enzymes may optionally originate from the non-animal organism, may be added and / or may be present in the form of microorganisms and / or secreted from such. In one embodiment, fermenting may be used for increasing umami taste. In one embodiment, fermenting may be used for increasing shelf-life. In one embodiment, a fermented comestible nutrient composition may, after the step of fermenting, comprise at least 0.5% by weight, at least 1 % by weight, at least 2% by weight, at least 5% by weight, at least 10% by weight, based on the total weight of the comestible nutrient composition.

[2153] A step of thickening may be conducted by any means. It may comprise partial removal of water and / or addition of further ingredient having thickening properties such as, e.g. one or more sugars, one or more other (poly)saccharides (e.g. starch, resistant starch, agar, pectin, inulin, fructans, raffinose, polydextrose), one or more thickening sweeteners, one or more thickening agents (e.g., cellulose, hemicellulose, carboxymethyl cellulose, carboxy ethyl cellulose, carboxypropyl cellulose, chitin, beta-glucan, raw guar gum, xanthan gum, lignin, a polyuronide, alginic acid or a salt thereof (e.g., sodium alginate), or a combination of two or more thereof) or a combination of two or more thereof.

[2154] A step of coagulating the collagen, optionally comprised in a comestible nutrient composition, may be conducted by any means. For instance, it may be conducted by adding one or more components facilitating coagulation such as, e.g., one or more collagens (e.g., one or more enzymes, preferably one or more (poly)peptidases / proteases (e.g., chymosin, pepsin) and / or one or more lipases, in particular enzyme mixtures such as, e.g., lab (also: rennet)), acidification, heating, cooling, one or more freeze / thaw cycles, or a combination of two or more thereof. Enzymes used for this purpose, including lab, may be obtained from any natural sources or from any biotechnological means. This may optionally provide a cheeselike semi-solid product (“curd”) and separate it from a liquid phase (“whey”). For instance, casein may be coagulated by addition of one or more enzymes (e.g., lab). This may allow preparing a curd containing casein and a whey fraction containing the non-coagulated components. This may allow preparing a cheese-like product from casein. Herein, the casein and / or the one or more enzymes used for coagulation may be obtained as the collagen from the method of the present invention. Optionally, such comestible nutrient composition may form a dried powder. If so, such dried powder may be the final product (comestible nutrient composition). Alternatively or additionally, such powder may be re-dissolved or resuspended in a suitable liquid such as, e.g, water, a consumable aqueous buffer, or a mixture of one thereof comprising ethanol. Accordingly, on a preferred embodiment, the comestible nutrient composition is a drinkable composition, and the method includes the further step of suspending the powder in an aqueous liquid, in particular in mineral or tap water. Alternatively or additionally, such powder may be further processed into a granulate.

[2155] In one embodiment, the comestible nutrient composition is:

[2156] (a) a drinkable composition such as a drink;

[2157] (b) a powder or granulate composition;

[2158] (c) a gel; or

[2159] (d) a frozen or partly frozen composition, wherein the comestible nutrient composition may optionally form part of a filling of a capsule or may optionally form part of a drink, a dairy product, a non-dairy cream, a sauce, or a bakery good.

[2160] The collagen and optionally further parts of the non-animal organism may optionally form part of a drink or foodstuff or a gel.

[2161] As used herein, the terms “drink” and “beverage” may be understood interchangeably in the broadest sense as generally understood in the art as a liquid or syrup-like orally consumable composition. It may or may not contain alcohol. A drinkable composition may be in liquid or pasty form, preferably in liquid form.

[2162] Optionally, a lyophilized or dried product (which may be, e.g., a powder or foamlike) may be milled to form a homogeneous powder. Such optional milling process may be applied using a pulverizer.

[2163] As used herein, the term “gel” may be understood in the broadest sense and may also comprise a gel as such, any kind of jelly composition, a filling of a capsule, filling of a bakery good, a pudding form, etc.

[2164] The comestible nutrient composition may also be a frozen or partly frozen composition, including an ice cube, frozen yogurt-like, soft ice like, slushies and the like. A further aspect of the present invention refers to the use of a comestible nutrient composition of the present invention for providing a well-defined nutrient composition, in particular a well-defined protein composition, to a consumer.

[2165] The comestible nutrient composition may optionally further comprise one or more further consumable ingredients selected from the group consisting of one or more vitamins, one or more minerals, one or more aroma compounds, one or more food colors, one or more types of fibers, ethanol, acetic acid, carbonic acid, and combinations of two or more thereof. Ingredients may interchangeably also be designated as components, additives, etc.

[2166] This may allow compensating for common micronutrient deficiencies of consumers (e.g., iron, vitamin A and iodine, but possibly also zinc, folate, vitamin B12, other B vitamins, vitamin C, vitamin D, calcium, selenium and fluoride).

[2167] A vitamin may be any vitamin known in the art. For instance, vitamins that may be added are selected from the group consisting of vitamin A, vitamins B (e.g., vitamin B1 , vitamin B2, vitamin B3, vitamin B5, vitamin B6, folate, vitamin B12), vitamin C, vitamin D, and metabolic precursors and combinations of two or more thereof. A vitamin or metabolic precursor thereof may be of natural or synthetical origin (nature-identical in structure or artificial in structure). It may be commercially available.

[2168] Preferably, the comestible nutrient composition comprises not more than 20% by weight, not more than 10% by weight, not more than 5% by weight, not more than 2% by weight, or not more than 1 % by weight, referred to the total mass of the comestible nutrient composition, of total amount of vitamins.

[2169] A mineral may be any nutrient that may be of nutrient value for consumers. Typically and preferably, minerals comprise ions or complexes of metals such as, e.g., a metal selected from the group consisting of iron, copper, calcium, zinc, and a combination of two or more thereof. Furthermore, minerals may also be selenium, iodine, and / or fluoride. A mineral may be commercially available. Preferably, the comestible nutrient composition comprises not more than 20% by weight, not more than 10% by weight, not more than 5% by weight, not more than 2% by weight, or not more than 1 % by weight, referred to the total mass of the comestible nutrient composition, of total amount of minerals. An aroma compound may be any comestible compound known in the art that alters the flavor or taste. Preferably, an aroma compound has a significant impact on the comestible nutrient composition’s taste, when it is applied in low amounts. Preferably, the comestible nutrient composition comprises not more than 20% by weight, not more than 10% by weight, not more than 5% by weight, not more than 2% by weight, or not more than 1 % by weight, referred to the total mass of the comestible nutrient composition, of total amount of aroma compounds. An aroma compound may be of natural or synthetical origin (nature-identical in structure or artificial in structure). For example, an aroma compound may be menthol furaneolhexyl cinnamaldehyde, isovaleraldehyde anisic aldehyde cuminaldehyde, glutamate, fructone, ethyl methylphenylglycidate, dihydrojasmone, oct-1 -en-3-one, 2-acetyl-1 -pyrroline, 6-acetyl-2,3,4,5-tetrahydropyridine, delta-octalactone, massoia lactone, diacetyl acetoin, nerolin, and combinations of two or more thereof. Many aroma compounds are known by those skilled in the art and may be commercially available.

[2170] A food color may be any comestible dye known in the art that alters the color of food, also designatable as food coloring. Preferably, a food color has a significant impact on the comestible nutrient composition’s color, when it is applied in low amounts. The coloring may be freely selectable.

[2171] Preferably, the comestible nutrient composition comprises not more than 20% by weight, not more than 10% by weight, not more than 5% by weight, not more than 2% by weight, or not more than 1 % by weight, referred to the total mass of the comestible nutrient composition, of total amount of food colors. A food color may be of natural or synthetical origin (nature identical in structure or artificial in structure). For example, a food color may be beetroot juice, beta-carotene, quinoline yellow, Ponceau 4R, Patent blue V, Green S, Brilliant blue FCF, Citrus red 2, Orange B Indigotine, Fast green FCF, Erythrosine, Allura red AC, Tartrazine, Sunset yellow FCF, or a combination of two or more thereof. A food color may even be fluorescent such as fluorescein. Many food colors are known by those skilled in the art and may be commercially available.

[2172] The comestible nutrient composition may have any texture, which may be defined by optionally added to the comestible nutrient composition. For example, as texturizing and / or nutrient, one or more further collagens of extracellular matrix (e.g. , collagen, elastin, etc.) may be added to the comestible nutrient composition. Fibers may be any comestible fibers, which may also be designated as dietary fibers. Typically, fibers in the context of the present invention are plant-derived food ingredients that cannot be completely broken down by human digestive enzymes. For example, it may originate from legumes, whole grains and cereals, vegetables, fruits, nuts or seeds. For example, fibers may comprise or consist of cellulose, hemicellulose, chitin, pectin, resistant starch, or a combination of two or more thereof. Fibers may also comprise inulin, beta-glucan, raw guar gum, xanthan gum, fructans, lignin, a polyuronide, an alginic acid or a salt thereof (e.g., sodium alginate), agar, carrageen, raffinose, polydextrose, or a combination of two or more thereof. Many fibers are known by those skilled in the art and may be commercially available.

[2173] Based on the production process of the present invention, but also as component of the admixed finished comestible, proteins of the extracellular matrix (ECM) may be comprised in the composition, such as, e.g., collagen type I, hyaluronic acid, polylysin, vinculin, laminin, fibronectin.

[2174] Optionally, the comestible nutrient composition may comprise one or more juices, optionally in addition to one or more of the further ingredients mentioned herein. For instance, it may comprise one or more fruit or vegetable juices.

[2175] The pH of the comestible nutrient composition may be adjusted to a desired range. Preferably, the pH is essentially neutral or sour. In one embodiment, the pH is in the range of pH 2-9, pH 2-8, pH 3-7, pH 4-7, pH 5-7, or pH 4-6. The pH may be adjusted by comestible acids, bases, buffer agents and / or acidity regulators.

[2176] In a preferred embodiment, a high-grade nutritional liquid product for individual demands may be obtained. The comestible nutrient composition may be adjusted to individual lifestyle demands, thus, its content ranges may be adapted to the desire of a specific consumer or group of consumers. This may be considered as “customizable food” and / or “personalized food”.

[2177] The present invention also refers to a method of preparing the collagen that naturally occurs extracellularly in one or more animal species. Thus, a further aspect of the present invention relates to a method for preparing a collagen that naturally occurs extracellularly in one or more animal species, comprising the steps of: (i) providing a non-animal organism comprising genetic information encoding for the collagen that naturally occurs extracellularly in one or more animal species;

[2178] (ii) expressing the collagen in the non-animal organism and optionally subjecting the collagen to one or more posttranslational modifications.

[2179] It will be understood that the definitions and preferred embodiments laid out in the context of the non-animal organism as laid out above mutatis mutandis apply to the method for preparing the collagen.

[2180] It will be understood that the process may be conducted in various size ranges. It may optionally be conducted in a laboratory scale such as, e.g, in a flask, or in a large industrial scale such as, e.g., in a fermenter. Likewise, the mass of non-animal organism may be adapted.

[2181] The conditions may be adapted to the non-animal organism chosen. For example, some organisms may be supported by adding oxygen, others may prefer the essential absence of gaseous oxygen. Likewise, the temperature may be adapted to each organism. A temperature range at which the non-animal organism expresses a collagen may be chosen depending on the conditions. In certain embodiments, it is in the range of 18 to 40°C, 18 to 22°C, 22 to 30°C, or 35 to 38°C.

[2182] Providing the non-animal organism may be conducted by any means. The non- animal organism may be optionally directly prepared such as, e.g., by transfecting the non-animal organism with genetic material encoding for the collagen. Alternatively, it may be a permanently genetically modified organism that may optionally also be breeded. Optionally, the non-animal organism may be obtained from a stock and may optionally be (deep)frozen.

[2183] Expressing the collagen may be performed permanently or may be induced in case of using an inducible promoter and / or enhancer. The collagen expression may also be transient in case of using plant hosts (e.g. N. tabacum, N. benthamiana)

[2184] In a preferred embodiment, the method further comprises one or more of the following: harvesting the non-animal organism that has expressed the collagen; isolating the collagen from the non-animal organism or cell-culture medium in which the non-animal organism is cultivated; preserving the collagen or composition comprising such from spoiling; and / or freeze-drying or drying of the collagen or composition comprising such.

[2185] As used herein, the term “harvesting” may be understood in the broadest sense as any means for obtaining the non-animal organism. For instance, a monocellular organism such as, e.g., bacterium or yeast, may be harvested by centrifugation and optional washing steps. For instance, a multicellular organism such as, e.g., a plant or a fungus, may be harvested by chopping or picking the parts of interest. The parts of interest of a plant may exemplarily be the aerial parts or the roots, the leaves, the stem, the blossoms, the flowers, and / or the fruits. Harvesting may include one or more steps of releasing material therefrom and optionally lysing cells. Thus, harvesting may include exposing of cells of the non-animal organism, parts of the non-animal organism, or the non-animal organism as a whole to a lysis buffer (e.g., containing one or more surfactants) and / or sonication. This may optionally be combined with one or more further centrifugation steps.

[2186] As noted above, the collagen may be located inside the non-animal organism or may be located outside. The collagen may be isolated from the non-animal organism or the surrounding environment such as the cell culture medium in which the non- animal organism is cultivated. As used herein, the term “isolating” may be understood in the broadest sense as obtaining the collagen and separating it from other parts of the non-animal organism or the surrounding environment. It does not have to be entirely pure. Isolating may also comprise a step of precipitation such as, e.g, by addition of salt (salting out, e.g., by means of ammonium chloride, such as 20 to 70%, or 40 to 60% of ammonium chloride), or addition of an anti-solvent (e.g. ethanol or other organic solvents), or reducing the salt concentration (salting in). The person skilled in the art will be aware of means and will be able to adapt these to the respective collagen to be purified.

[2187] Optionally, isolating may also include the coagulation. Coagulation may be conducted by any means such as, e.g., adding one or more components facilitating coagulation such as, e.g., one or more collagens (e.g., one or more enzymes, (e.g., one or more (poly)peptidases / proteases such as, e.g., chymosin, pepsin) and / or one or more lipases), acidification, heating, cooling, one or more freeze / thaw cycles, or a combination of two or more thereof. Coagulation may separate a fraction containing one or more coagulated components (curd-like fraction) and a fraction containing one or more components that remain soluble in the liquid fraction (wheylike fraction). Depending on the characteristics of the collagen of interest expressed by the non-animal organism, this may be located in the coagulated fraction or the liquid fraction. Optionally, the fraction of interest may be treated further by any means.

[2188] Optionally, isolating may also include the removal of salt such as, e.g., via dialysis and / or chromatographic means (e.g., size-exclusion chromatography (SEC)). Dialysis and / or chromatographic means (e.g., size-exclusion chromatography (SEC)) may optionally also be used for increasing the concentration of the collagen.

[2189] As used herein, the term “preserving” may be understood in the broadest sense as any means for preventing spoiling. For instance, one or more preserving agents may be added which may be preferably comestible preserving agents such as, e.g., benzoic acid. As indicated above, the collagen or a composition containing such may be subjected to fermenting, smoking, pickling and / or thickening to preserve. The collagen or a composition containing such may be optionally heated such as, e.g., pasteurized. The collagen or a composition containing such may optionally also be freeze-dried, frozen, dried, or cooled to preserve it.

[2190] The collagen or a composition containing such may optionally be subjected to freeze-drying or drying. This may be achieved by routine means. In a preferred embodiment, this is achieved by chromatographic means such as, e.g., affinity chromatography, size-exclusion chromatography (SEC), ion exchange chromatography (IEX), reverse-phase chromatography, high-performance liquid chromatography (HPLC), ultra high-performance liquid chromatography (LIPLC), fast protein liquid chromatograph (FPLC), or a combination of two or more thereof. Such chromatographic step may be conducted at any suitable temperature, preferably in a temperature range of 0 to 30°C, 3 to 15°C or 15 to 25°C. Optionally, the purity of the collagen may be determined.

[2191] An exemplified, non-limiting example flow scheme is shown in Figure 18A, where it will be understood that such process may be modified and is not limited to Pichia pastoris and not limited to collagen purification. In addition, an exemplified, nonlimiting example flow scheme depicting the complementary harvesting of POI from supernatant, is shown in Figure 18B.

[2192] A further exemplified, non-limiting example flow scheme is shown in Figure 19, where it will be understood that such process may be modified and is not limited to Pichia pastoris and not limited to collagen purification. A still further exemplified, non-limiting example flow scheme is shown in Figure 24, where it will be understood that such process may be modified and is not limited to plants and not limited to collagen purification.

[2193] A still further exemplified, non-limiting example flow scheme is shown in Figure 32, where it will be understood that such process may be modified and is not limited to Pichia pastoris and not limited to collagen purification.

[2194] A still further exemplified, non-limiting example flow scheme for upscaling the process is shown in Figure 37.

[2195] Further exemplified, non-limiting examples of detailed process schemes are shown in Figures 38 and 39.

[2196] A still further exemplified, non-limiting example flow scheme for preparing collagen from plants is shown in Figure 40, where it will be understood that such process may be modified and is not limited to plants.

[2197] A still further exemplified, non-limiting examples of detailed process schemes is shown in Figure 41 .

[2198] Optionally, the purity of the collagen may be further improved. In a preferred embodiment, the method comprises purification of the collagen by affinity chromatography. For example, the content of the collagen may be at least 1 % by weight, at least 5% by weight, at least 10% by weight, at least 25% by weight, at least 50% by weight, at least 70% by weight, at least 80% by weight, or at least 90% by weight, based on the total (poly)peptide / collagen content.

[2199] Affinity chromatography may be performed by any means. As indicated above, affinity chromatography may be based on a tag such as, e.g., a His-tag that is selectively bound by a binding partner, in case of a His-tag, for example, nickel ions. An affinity chromatography column may be loaded with the collagen, which may be eluted by using an elution agent such as, e.g., in case of a His-tag, a nickel-binding agent that may disturb the binding of the His-tag to the column.

[2200] In a preferred embodiment, the method further comprises:

[2201] (iii) preparing a solution of the collagen in dissolved form, optionally filtering and / or centrifugation of the solution to remove any remaining solid particles or debris and obtaining a clear collagen-containing solution, and optionally precipitating the collagen;

[2202] (iv) isolating the collagen of step (iii) via chromatography and collecting collagen- containing fractions;

[2203] (v) optionally subjecting the collagen of step (iv) to dialysis or buffer exchange via a size exclusion chromatography column.

[2204] In a preferred embodiment, the method further comprises isolating the collagen by affinity chromatography, preferably by an antibody and / or by binding to a tag attached to the collagen, in particular a His-tag.

[2205] The obtained collagen may be stored at any conditions suitable for such storage. For instance, the collagen may be stored in solution or in dry state (e.g., dried or freeze-dried). It may be stored at any suitable temperature such as, e.g., in a range of -100 to -20°C, -20 to 0°C, 1 to 10°C, or 10 to 30°C, for example in liquid nitrogen, in a deep freezer (e.g., -90°C to -70°C), in a freezer (e.g., -25 to -5°C), in a fridge (e.g., 2 to 8°C) or at ambient temperature (e.g., 18 to 25°C).

[2206] As indicated above, it is of interest to prepare a comestible nutrient product. Thus, a further aspect of the present invention relates to a method for preparing a comestible nutrient product, in particular a vegetarian comestible nutrient product, comprising the step of adding the collagen that naturally occurs extracellularly in one or more animal species obtained from a method of preparing a collagen of the present invention to one or more further comestible nutrient ingredients.

[2207] It will be understood that the definitions and preferred embodiments laid out in the context of the non-animal organism and the method for preparing the collagen as laid out above mutatis mutandis apply to the method for preparing a comestible nutrient product.

[2208] As indicated above, such comestible nutrient product may be any comestible nutrient product known in the art such as, e.g., a beverage, a drink, food, etc.

[2209] As indicated above, the present invention also comprises a comestible nutrient product obtainable from the methods of the present invention. Thus, a further aspect refers to a comestible nutrient product, in particular a vegetarian comestible nutrient product, comprising at least one collagen that naturally occurs extracellularly in one or more animal species obtained from a method of the present invention. It will be understood that the definitions and preferred embodiments laid out in the context of the non-animal organism, the method for preparing the collagen and the method for preparing a comestible nutrient product as laid out above mutatis mutandis apply to the comestible nutrient product.

[2210] The following Examples as well as the accompanying Figures and claims are intended to provide illustrative embodiments of the present invention described and claimed herein. These Examples and Figures are not intended to provide any limitation on the scope of the invented subject-matter.

[2211] Brief Description of the Figures

[2212] Figure 1 shows His-tag purification of Collagen from P. pastoris (20 ml), wherein the Y-axis shows conductivity in mS / cm, the UV detection at 280 nm (1 ), the conductivity (2) and the gradient of eluent (3) is depicted.

[2213] Figure 2 shows mass spectrometry results of detected peptides aligned to the sequence of recombinant collagen. The black bars indicate hydroxylated amino acid residues. Besides proline hydroxylation, there is also a methionine hydroxylation that occurs during the preparation of samples. Methionine hydroxylation appears more frequently with much larger bars. His-tag was not detected.

[2214] Figure 3 shows an agarose gel analysis of the Pichia pastoris clones of Example 1 . The lanes are as follows: 10 kb ladder (1 ), Clones A1 -A8 (2-9), 100 bp ladder (10), empty (11 and 13), negative control (12), and positive control (14).

[2215] Figure 4 shows the regeneration efficiency of Nicotiana benthamiana dependent on the concentration of selection agent (BASTA) and the Agrobacterium strain (EHA105, GV3101 and LBA4404) used for transformation. Regeneration Media (Musashiagei-Skoog media (MSII)) including Timentin, Terbinafin, Cefotaxime (MSII-TTC). From left to right, the results depict BASTA concentration of 0, 0.5, 1 .0, 1 .5, 2.0, 3.0 and 4.0 mg / ml. The Y-axis shows the number of regenerated explants. Regenerated explants have been scored wild-type (WT), if green, chimera, if not fully red and RUBY, if entire explant was red.

[2216] Figure 5 shows the regeneration efficiency of Nicotiana benthamiana dependent on the concentration of selection agent (BASTA) and the Agrobacterium strain (EHA105, GV3101 and LBA4404) used for transformation. Regeneration Media (Musashiagei-Skoog media (MSII)) including NAA and BAP (1 -Naphthaleneacetic acid (NAA), 6-Benzylaminopurin (BAP) (MSII-NB)). From left to right, the results depict BASTA concentration of 0, 0.5, 1 .0, 1 .5, 2.0, 3.0 and 4.0 mg / ml. The Y-axis shows the number of regenerated explants. Regenerated explants have been scored wild-type (WT), if green, chimera, if not fully red and RUBY, if entire explant was red.

[2217] Figure 6 shows the scheme of rheology measurements. The top plate oscillates at a given stress or strain amplitude. In the present context, it preferably is set to a present strain amplitude.

[2218] Figure 7 shows the determination of rheology measurements. The measurements provide a gelation experiment with commercial PureCol 5 mg / mL at 37°C (triplicate). The results are given in stirage modulus G’ in Pa (shear strain y [%] = 3; angular frequency w [1 / s] = 10).

[2219] Figure 8 shows a native polyacrylate gel electrophoresis (PAGE) gel, testing Collagen expression in / V. benthamiana. Herein: COL- = Coll a, COL+ = Coll a + P19, COLa = Colla + P4H-a, COLb = Colla + P4H-B, and COLab = Colla + P4H- a + P4H-B.

[2220] Figure 9 shows immuno-blotted polyacrylate gel electrophoresis (PAGE) gels with collagen with / without P4H-a / P4H-b expression in N. benthamiana at different time points as depicted. Herein: COL- = Colla, COL+ = Coll a + P19, COLa = Coll a + P4H-a, COLb = Coll a + P4H-B, and COLab = Coll a + P4H-a + P4H-B.

[2221] Figure 10 shows a stainfree PAGE 4-20% gel with samples of different time points of Collagen expression in N. benthamiana.

[2222] Figure 11 shows an immunoblot with x6His with samples of Collagen expression in N. benthamiana at different time points.

[2223] Figure 12 shows an SDS-PAGE 7,5% Coomassie staining of samples of Collagen expression in N. benthamiana at different time points.

[2224] Figure 13 shows an immunoblot with samples of different time points corresponding to Figure 12. Figure 14 shows an SDS PAGE of a collagen purification process from P. pastoris. The lanes are peqGOLD protein marker IV (1 ), PureCol 3 mg / ml (2), lysate (3), fraction A3-B11 (FT) (4), fraction A3-B2(FT) (5), fractions / peaks D4-D5 (6), fractions / peaks D6-D7 (7), and fractions / peaks D8-D9 (8).

[2225] Figure 15 shows the Western Blot of the collagen purification process of Figure 14. The lanes are peqGOLD protein marker IV (1 ), PureCol 3 mg / ml (2), lysate (3), fraction A3-B11 (FT) (4), fraction A3-B2(FT) (5), fractions / peaks D4-D5 (6), fractions / peaks D6-D7 (7), fractions / peaks D8-D9 (8), and negative control albumin (9).

[2226] Figure 16 shows a gel electrophoresis of purified collagen from Pichia pastoris. The lanes are peqGOLD protein marker IV (1 ), PureCol 3 mg / ml (positive control) (2), PanCol 1 mg / ml (positive control) (3), fraction D7 purified with His-tag (4), fractions D8-D11 purified with His-tag (5), fraction D11 purified with His-tag (6), fraction D1 purified with His-tag (7), fraction D2 purified with His-tag (8), fraction D3 purified with His-tag (9), and pooled fractions D7-D10 and rebuffered. The box and arrow indicate the sample which was investigated by mass spectrometry.

[2227] Figure 17 shows a Western Blot of a collagen purification process after P. pastoris culture optimisation steps. The lanes are peqGOLD protein marker IV (1 ), FFGF2 (2), albumin pellet (3), albumin SN (4), A8 pellet 20°C (5), A8 SN 20°C (6), A8 pellet 28°C (7), A8 SN 28°C (8), A11 pellet 20°C (9), A11 SN 20°C (10), A11 pellet 28°C (11 ), and A11 SN 28°C (12).

[2228] Figure 18A shows a general downstream processing flow scheme exemplified for collagen obtained from Pichia pastoris. “§” indicates that the process is pursued. Process steps are black boxes. Dashed boxes are optional steps and white solid boxes indicate analytical assays.

[2229] Figure 18B shows a general downstream processing flow scheme exemplified for collagen obtained from supernatant of Pichia pastoris, thus collecting secreted collagen. “§” indicates that the process is pursued. Frames indicate further analysed fractions.

[2230] Figure 19 shows a general downstream processing flow scheme exemplified for collagen from plants. “§” indicates that the process is pursued. Figure 20A shows an SDS PAGE of collagen obtained from the supernatant of Pichia pastoris. Captions 1 , 2 and 3 indicate different batches of supernatant (22 mL, 22 mL, 300 mL).

[2231] Figure 20B shows a Western Blot of collagen obtained from the supernatant of Pichia pastoris. Captions 1 , 2 and 3 indicate different batches of supernatant (22 mL, 22 mL, 300 mL); +- Pancol 0.2 mg / mL; Neg. - Pichia Albumin Supernatant (negative control).

[2232] Figure 21 A shows an SDS PAGE of collagen obtained from Pichia pastoris.

[2233] Figure 21 B shows a Western Blot of collagen obtained from Pichia pastoris.

[2234] Figure 22 shows a size exclusion chromatography (SEC) chromatogram of collagen obtained from Pichia pastoris, including a UV peak (1 ) and a conductivity peak (3). The concentration of additional agents was kept low.

[2235] Figure 23 shows an SDS PAGE of several fractions of the collagen peaks obtained from the SEC as shown in Figure 22.

[2236] Figure 24 shows a general downstream processing flow scheme exemplified for collagen obtained from plants. “§” indicates that the process is pursued.

[2237] Figure 25 shows a Western blot of purified collagen by using 6.8 g of plant material.

[2238] Figure 26 shows an SDS PAGE of collagen using 6.8 g of plant material. The lanes are peqGOLD protein marker IV (1), ProColl 1.5 mg / ml (2), fraction V1-10 1 :5 (3), fraction V1-10 40% ammonium sulfate (AS) precipitation (4), fraction V1-10 60% ammonium sulfate (AS) precipitation (5), fraction V19 1 :5 (6), fraction V19 40% ammonium sulfate (AS) precipitation (7), and fraction V19 60% ammonium sulfate (AS) precipitation (8).

[2239] Figure 27 shows a Western blot of purified collagen by using 1 g of plant material. The lanes are peqGOLD protein marker IV (1 ), ProColl (2), Tris 1 :10 (3), Tris 40% ammonium sulfate (AS) precipitation (4), Tris 60% ammonium sulfate (AS) precipitation (5), Tris 1 :10 negative (6), Tris 40% ammonium sulfate (AS) negative (7), Tris 60% ammonium sulfate (AS) negative (8), and acetic acid SN negative (9). Figure 28 shows an SDS PAGE of collagen using 6.8 g of plant material. The lanes are peqGOLD protein marker IV (1 ), ProColl (2), Tris 1 :10 (3), Tris 40% ammonium sulfate (AS) precipitation (4), Tris 60% ammonium sulfate (AS) precipitation (5), Tris 1 :10 negative (6), Tris 40% ammonium sulfate (AS) negative (7), Tris 60% ammonium sulfate (AS) negative (8), and acetic acid SN negative (9).

[2240] Figure 29 shows a Western blot of purified collagen by using 5.8 g of plant material with polypeptide concentration of ca. 7 mg / ml on the gel. The lanes are peqGOLD protein marker IV (1 ), ProColl 0.25 mg / ml (2), Tris 16 (3), 40% ammonium sulfate (AS) precipitation 16 (4), 60% ammonium sulfate (AS) precipitation 16 (5), Tris 6 (6), 40% ammonium sulfate (AS) negative (7), 60% ammonium sulfate (AS) negative (8), sample buffer (9), and sample buffer (10).

[2241] Figure 30 shows a Western blot of purified collagen by using 7.5 g of plant material. The lanes are peqGOLD protein marker IV (1 ), ProColl 1.5 mg / ml (2), V1 -3 acetic acid (AA) 1 :3 (3), V1 -3 0.4 M (4), V1 -3 0.9 M (5), V1 -3 SN (6), 1 -20 AA 1 :3 (7), V1 - 20 0.4 M (8), V1 -20 0.9 M (9), and V1-20 SN (10).

[2242] Figure 31 shows an SDS PAGE of collagen using 6.8 g of plant material. The lanes are peqGOLD protein marker IV (1 ), ProColl 1.5 mg / ml (2), V1 -3 acetic acid (AA) 1 :3 (3), V1 -3 0.4 M (4), V1 -3 0.9 M (5), V1 -3 SN (6), 1 -20 AA 1 :3 (7), V1 -20 0.4 M (8), V1 -20 0.9 M (9), and V1 -20 SN (10).

[2243] Figure 32 shows a general downstream processing flow scheme exemplified for proteins purified from Pichia pastoris.

[2244] Figure 33 shows an SDS PAGE of collagen obtained from Pichia pastoris. The lanes are peqGOLD protein marker VI (1 ), Pichia A2, Lysate after 100 h (2), Pichia A3, Lysate after 100 h (3), Pichia A4, Lysate after 100 h (4), Pichia WT Negative Control, Lysate after 100 h (5), Pichia A8, Lysate after 72 h (6), Pichia A9, Lysate after 72 h (7), Pichia A10, Lysate after 72 h (8), Pichia A11 , Lysate after 72 h (9), and PureColl 0.3 mg / ml (10).

[2245] Figure 34 shows a SDS PAGE and Western Blot (fraction) of collagen obtained from Pichia pastoris. The lanes are peqGOLD protein marker VI (1 ), PanCol 0.25 mg / ml (2), Tris 16 (3), 40% ammonium sulfate (AS) (4), AS60% 16, (5), Tris 6 (6), AS40% 6 (7), AS60% 6 (8), sample buffer (9), sample buffer (10), Pichia albumin control (11 ), Pichia A8 (12), and Pichia AH (13). Lower part of the picture shows the bands after a longer exposure time.

[2246] Figure 35 shows a gel electrophoresis of E. coli colonies. The lanes are 10 kB ladder (1 ), clones 1 -6 (2-7), clone 8 (8), clone 9 (9), 100 bp ladder (10), negative control (11 ), and 10 kb ladder (12).

[2247] Figure 36 shows SDS PAGE (above) and Western Blot (below) of collagen obtained from E. coli. The lanes are peqGOLD protein marker IV (1 ), PureCol positive control (2), PanCol positive control (3), Negative control spiked with 0.75 mg / ml ProCol (4), extraction buffer (5 and 6), extraction buffer negative (7 and 8), E. coli (9 and 10).

[2248] Figure 37 shows an upscaling flow scheme exemplified for preparing collagens. Reference: de Sa Magalhaes, S.; Keshavarz-Moore, E. Pichia pastoris (Komagataella phaffii) as a Cost-Effective Tool for Vaccine Production for Low- and Middle-Income Countries (LMICs). Bioengineering 2021 , 8, 119. https: / / doi.Org / 10.3390 / bioengineering8090119.

[2249] Figure 38 shows an example of a detailed process scheme for preparing collagens. Reference: da Silva Ferreira, Adamo Eduardo: Simulation and techno-economic analysis of production processes of novel generation L-Asparaginases using recombinant Escherichia coli and Pichia pastoris. Dissertagao (Mestrado) - Escola Politecnica da Universidade de Sao Paulo. Departamento de Engenharia Quimica Sao Paulo, 2022. https: / / www.teses.usp.br / teses / disponiveis / 3 / 3137 / tde- 23032023-071933 / publico / AdamoEduardodaSilvaFerreiraCorr22.pdf.

[2250] Figure 39 shows an example of a detailed process scheme for preparing collagens. Reference: Nandi, Sowen, Aaron T. Kwong, Barry R. Holtz, Robert L. Erwin, Sylvain Marcel & Karen A. McDonald (2016) Techno-economic analysis of a transient plantbased platform for monoclonal antibody production, mAbs, 8:8, 1456-1466, DOI: 10.1080 / 19420862.2016.1227901 .

[2251] Figure 40 shows an example of a flow scheme for preparing collagens from plants. Reference: Park, Se-Ra & Lim, Chae-Yeon & Kim, Deuk-Su & Ko, Kisung. (2015). Optimization of Ammonium Sulfate Concentration for Purification of Colorectal Cancer Vaccine Candidate Recombinant Protein GA733-FcK Isolated from Plants. Frontiers in Plant Science. 6. 10.3389 / fpls.2015.01040. Figure 41 shows an example of a detailed process scheme for preparing collagens from plants. Reference: Alam A, Jiang L, Kittleson GA, Steadman KD, Nandi S, Fuqua JL, Palmer KE, Tuse D, McDonald KA. Technoeconomic Modeling of Plant- Based Griffithsin Manufacturing. Front Bioeng Biotechnol. 2018 Jul 24;6:102. doi: 10.3389 / fbioe.2018.00102. PMID: 30087892; PMCID: PMC6066545.

[2252] Figure 42 shows vector map of vector Col1a_1 (35S_H is_ full length).

[2253] Figure 43 shows vector map of vector Col1a_2 (35S_His_ full length_no tag).

[2254] Figure 44 shows vector map of vector Col1a_7 (P19_pPD7-His-TEV-Col1a(triple helical domain only)-AFVY-tNos).

[2255] Figure 45 shows vector map of vector Col1a_8 (P19_pPD7-His-TEV-Col1a(triple helical domain only)-AFVY-tNos).

[2256] Figure 46 shows vector map of vector Col1a_9 (P19_pPD7-fuGFP-TEV- Col1 a(triple helical domain only)-AFVY-tNos).

[2257] Figure 47 shows vector map of vector Col1a_10 (P19_pPD7-fuGFP-TEV- Col1 a(triple helical domain only)-tNos).

[2258] Figure 48 shows vector map of vector Col1a_11 (P19_pPD7-fuGFP-TEV- Col1 a(triple helical domain only)-KDEL-tNos).

[2259] Figure 49 shows vector map of vector P4Ha_1 .

[2260] Figure 50 shows vector map of vector P4Hbeta_1 .

[2261] Figure 51 shows vector map of Coll A1 expression vector for E.coli.

[2262] Figure 52 shows vector map of Coll A1 expression vector for Pichia pastoris.

[2263] Examples

[2264] Example 1 - Transformation of Pichia pastoris with Col1A1 The aim of the experiment is to transform Pichia pastoris with the Col1A1 gene, without N- and C- terminal propeptides, in order to produce recombinant Collagen.

[2265] Methods

[2266] Transformation:

[2267] Transformation of competent Pichia pastoris was achieved by electroporation with freshly made competent cells. Following plasmid has been used: pPP-Zeocin-AOX1 >{Col1A1 without propept}

[2268] Plasmid has arrived in E. coli (Vectorbuilder), and before the transformation the plasmids were extracted with a MaxiPrep Kit (Qiagen).

[2269] The linearization of plasmid was carried out with a Pmel endonuclease (New England Biolabs) according to the manufacturer's manual. Linearization was verified by agarose gel electrophoresis. Ca. 2 pg of plasmid were used for transformation. The incubation time after the electroporation was 2 h 20 min. The incubation on YPDS agar plates with 200 pl of 1 and 2 mg / ml Zeocin spread over agar took 3 days. As a negative control one aliquot was electroporated without addition of plasmids.

[2270] PCR Screening:

[2271] For the colony, PCR clones were grown in 3 ml YPD medium with 100 pg / ml Zeocin overnight, at room temperature and 220 rpm, in 15 ml tubes. After spinning down 1 ml in 1.5 ml tubes 500 pl of the lysis buffer (400 mM TRIS-HCI, pH 8; 60 mM EDTA, pH 8; 150 mM NaCI, 1 % SDS) was added together with a spatula tip of 0.25 glass beads. The samples were disrupted for 5 min in the bead beater at 30 Hz and incubated for 10 minutes. After that 150 pl of 3 M Potassium Acetate was added and the tubes were vortexed briefly before centrifugation (1 min 14.000 g). Supernatants were transferred in a fresh tube and mixed with same amount of Isoporopanol. After another centrifugation for 5 min at 14.000 g the pellets were washed with 500 pl 70% Ethanol and dried before the dissolution in 100 pl MilliQ H2O.

[2272] Primers: 73 GACTGGTTCCAATTGACAAGC (SEQ ID NO: 35) 74 GCAAATGGCATTCTGACATCC (SEQ ID NO: 36) Expected band sizes for Coll A1 ca. 3200 bp PCR Programm: 33 Cycles 98°C 30 sec

[2273] 98°C 5 sec

[2274] 61 °C 5 sec

[2275] 72°C 45 Sec 72°C 1 min

[2276] 4°C

[2277] The Colony PCR results are shown in Figure 3. The lower bands show the native AOX1 from Pichia. The result is not conclusive due to a band in the negative control, possibly because of cross-contamination. Previous screenings have shown that most of the colonies that have grown on Zeocin contain the target gene.

[2278] Conclusion

[2279] The transformations were successful. So many clones were generated, that it would make sense to dilute the cells before plating them out or increase the antibiotic concentration. PCR Screening can be repeated or the colonies can be screened directly for the protein expression.

[2280] The clones are transferred on master plates and screened for production of collagen in liquid culture.

[2281] Example 2 - Example of stable transformation of plants

[2282] Stable transformation of plants is a technique of interest in plant biotechnology. It allows scientists to introduce new genes into plants, thereby altering their characteristics or conferring resistance to pests, diseases, or environmental stressors:

[2283] 1 . Choice and Preparation of Explant Tissue: o The process begins by selecting an appropriate tissue sample (explant) from the plant. Common explants include leaf segments, stem sections, or immature embryos. o These explants are then cultured in a nutrient-rich medium to encourage cell division and regeneration.

[2284] 2. DNA Delivery: o The next step involves introducing the desired DNA (transgene) into the plant cells. There are various methods for DNA delivery:

[2285] ■ Agrobacterium-Mediated Transformation: Agrobacterium tumefaciens, a soil bacterium, is used to transfer the transgene into the plant cells. This method is widely employed for many plant species.

[2286] ■ Particle Bombardment (Gene Gun): Tiny gold or tungsten particles coated with DNA are shot into the plant tissue using a gene gun. This method is particularly useful for species that are less amenable to Agrobacterium transformation.

[2287] ■ Other methods include electroporation and microinjection.

[2288] 3. Callus Induction and Regeneration: o After DNA delivery, the transformed cells are cultured on a special medium to form a mass of undifferentiated cells called callus. o The callus is then induced to differentiate into shoots and roots, ultimately leading to the development of whole plants. o Selection markers (such as antibiotic resistance genes) are often used to identify and propagate only those cells that have successfully incorporated the transgene.

[2289] 4. Recovery of Stably Transformed Plants: o Once the regenerated plants are established, they are transferred to soil and grown to maturity. o These stably transformed plants carry the introduced transgene in their genome and can pass it on to their offspring.

[2290] 5. Applications: o Stable transformation is used for various purposes:

[2291] ■ Crop Improvement: Developing genetically modified (GM) crops with enhanced traits (e.g., pest resistance, drought tolerance, improved nutritional content).

[2292] ■ Functional Genomics: Studying gene function by overexpressing or silencing specific genes.

[2293] ■ Biopharmaceutical Production: Producing therapeutic proteins in plants.

[2294] ■ Basic Research: Investigating plant biology and development. Remember, this process is essential for advancing our understanding of plant genetics and for developing sustainable agricultural solutions.

[2295] Detailed protocols are known, including insights into the steps involved

[2296] Example 3 - Nicotiana benthamiana Tissue Culture RUBY expression

[2297] Determine the most efficient conditions for tobacco tissue culture as an estimate for stable transformation of POI. Method:

[2298] Transformation:

[2299] Agrobacterium tumefaciens (A. tumefaciens', GV3101 ) strain carrying the transformation construct (35S-RUBY) inoculated in 50 ml culture induction medium (containing antibiotics), shake over night at 28°C. Day of transformation: Pellet bacteria by centrifugation, resuspend bacteria in MMA medium (without antibiotics) to an OD600=1 .5 (50 ml).

[2300] Harvest the 2-3 youngest, but fully expanded leaves of 4 weeks-old Nicotiana benthamiana plants.

[2301] Surface sterilize in 1 .2% NaOCI (+0.01 % Tween) and wash in H2O. Cut surface- sterilized leaves into pieces and in A. tumefaciens suspension. Incubate at least 30 min. Transfer leaf cuts onto water wetted paper. Seal dishes and incubate for 2 days in the dark at 24°C.

[2302] Selection and shoot induction

[2303] Prepare petri dish with 50 ml water (containing Cefotaxime (250 mg / l)) and place leaf cuts in it for washing. Gently shake the petri dish, and incubate for 10 min. Dry leaf cuts and place on shoot induction medium (MS-II). Use standard round petri dishes to minimize the risk of contamination. Place 8-10 leaf cuts on each plate. Incubate in a light cabinet until shoots occur at 25°C, 24 h.

[2304] Root induction

[2305] Shoots start developing from calli on MS-II plates and need to be transferred to MS- Ill plates to induce rooting. Cut well-developed shoots with a sterile blade. Stick shoots with the cut surface into MS-Ill medium, and incubate under the same conditions as before for further development of the shoot and rooting. Shoots can further develop in MS-Ill media, and can eventually form roots. Transfer well- developed shoots to soil.

[2306] Plant maturing and seed harvesting

[2307] Wait until the pods are dry and seeds get exposed before harvesting. Make a BASTA seeding selection / germination test in 1 / 2 MS with 1 mg / L Basta. Quantify the germination ratio.

[2308] Select survival seedlings and transfer them to pods for further generation selection. Quantification of the total number of shoots and its classification showed that overall Agrobacterium strain LBA4404 and Agrobacterium strain GV3101 were the most efficient strains for stable transformation as plants infected with both strains showed a higher number of chimeric shoots. However, GV3101 is clearly superior as it induced more chimeras per time interval.

[2309] Results

[2310] Results of the transformation efficiency in Murashige-Skoog media II (MSII) Timentin, Terbinafin, Cefotaxime (TTC) are depicted in Figure 4. Results of the transformation efficiency in Murashige-Skoog media II NAA, BAP (1- Naphthaleneacetic acid (NAA), 6-Benzylaminopurin (BAP) (MSII-NB)) are depicted in Figure 5.

[2311] The RUBY plant had red / purple flowers, while the wild-type had white flowers. The chimeric flowers were pink.

[2312] Example 4 - Transient expression of Collagen in Nicotiana benthamiana

[2313] Transient expression of Collagenl al from Bos taurus in N. benthamiana. All steps from infiltration to WB results are depicted below.

[2314] Method:

[2315] Transient Transformation:

[2316] Grow N. benthamiana plants for 4-5 weeks on long day conditions as: 16 h light (50% light and 60% humidity) / 8 h dark (0% light and 65% humidity). GV3101 Agrobacterium tumefaciens may be used for transient expression. Antibiotic selection for bacterial transformation is carried out according to the manufacturer.

[2317] Selected Agrobacterium strains carrying the genes Collagen 1 a, Hydroxylase P4H- a, Hydroxylase P4H-b were selected to further steps. Grow Agrobacterium in 2 mL of LB broth supplemented with the selection antibiotics in a test tube at 28°C for 1 day. Inoculate 500 pL of the preculture in 50 mL of LB with selection antibiotics and grow at 28°C over night.

[2318] Next day, wash cells 1x: Pellet at low speed (max. 3.000 g) and resuspend with 40 ml MMA (10 mM MgCI2, 10 mM MES [pH 5.7], 100 pM acetosyringone). Pellet again at low speed (max. 3.000 g) and resuspend with MMA (10 mM MgCI2, 10 mM MES [pH 5.7], 100 pM acetosyringone) media, adjust to final OD600 = 0.8.

[2319] Infiltrate 100 pL approx, in the leave with a needleless syringe following the next co infiltration patterns: 1. Colla

[2320] 2. Coll a + P4H-a

[2321] 3. Coll a + P4H-B

[2322] 4. Coll a + P4H-a + P4H-B

[2323] Harvest 2 to 4 days after infiltration.

[2324] Western blotting:

[2325] Harvest plant material from the marked infiltrated areas in the tobacco leaves into conical tubes.

[2326] Freeze the tissue in liquid nitrogen and grind the leaves to a fine powder while the plant material is still frozen. Add 400 pL of ice-cold extraction buffer:

[2327] Reagent Concentration

[2328] Tris / HCI (pH 8,5) 100 mM

[2329] Glycerol 10%

[2330] NaCI 0.5 M

[2331] Imidazole 20 mM

[2332] PVPP 5%

[2333] DTT 5 mM

[2334] Vortex vigorously to extract the soluble proteins; Centrifuge for 30 minutes at 18.000 RFC at 4°C Transfer the supernatant into a clean tube. Centrifuge once again for 20 minutes as in the previous step. Transfer the supernatant into a clean tube, keep sample in ice.

[2335] Total protein quantification via Bradford Assay according to manufacture:

[2336] Prepare samples to 15 pg, 30 pg or 50 pg (depending on the gel). Add H2O 4 x Loading Dye and DTT (1 :10). Denature samples for 5 min at 95°C in Thermomixer at 300 rpm. Cool samples on ice on the bench. Store at -20°C or proceed to electrophoresis.

[2337] Native PAGE gel

[2338] Dilute all the samples to the lower concentration and load the same amount of protein on each well. Using Bio-Rad stain free 15% polyacrylamide gel. Load 50 pL in the wells, flanking the samples with Chameleon duo color protein marker. Run the gel for 20 minutes at 80 V and then change the voltage up to 120 V until the samples reach the bottom of the gel. Dry blotting

[2339] According to iBIot™ 2 Gel TransferDevice manufacturer protocoll. Choose program P0 (20 V for 1 min, 23 V for 4 min, 25 V for 2 min) or program of choice.

[2340] Immunoblot

[2341] Prepare a 1 :1000 solution of primary antibody (His) and pour it into a box. Place the membrane into the box assuring that the antibody solution covers completely the membrane. Incubate the membrane with the primary antibody overnight with rocking motion. Discard the primary antibody solution and proceed to wash the membrane with TBS-T four times: 10 minutes, 5 minutes, 5 minutes, 10 minutes, Discard the TBS-T.

[2342] Prepare a 1 : 10000 solution with the secondary antibody. Incubate for 2 h at constant rocking motion. Discard the primary antibody solution and proceed to wash the membrane with TBS-T in four times: 10 minutes, 5 minutes, 5 minutes, 10 minutes, Discard the TBS-T.

[2343] Add 2 mL of reagents A and B on top of the membrane trying to cover all the surface. Use the transilluminator to record data.

[2344] Results:

[2345] Estimated time from seed to plant ready to infiltrate: 3.5 weeks. Double / triple construct infiltration shows more severe necrosis. Six leaf disks were sampled, two leaves per construct.

[2346] Table 3. Protein quantification. All data is presented in mg / mL.

[2347] The gel electrophoresis results (native gel and immuno-stained gel) are depicted in Figures 8 and 9.

[2348] Protein extraction modification:

[2349] Extraction buffer replaced with loading dye reducing SDS. Samples were boiled for 5 minutes. Three time points of harvesting after infiltration, all samples contain Col1 a+P19. Positive control (C+) is purified FgF2 9-point mutation (e.g., as described in EP 3380508). Negative control (C-) is protein extract from noninfiltrated leaves. SDS- PAGE stainfree 4-20%. The results are depicted in Figures 10 and 11.

[2350] Three time points of harvesting after infiltration, all samples contain Col1 a+P19. Positive control (C+) is purified FgF2 9 point mutation. Collagen coating solution (CC) is used as secondary positive control. Negative control (C-) is protein extract from non-infiltrated leaves. An SDS-PAGE 7,5% Coomassie staining was performed as depicted in Figure 12 as well as an immune-stain of a gel as shown in Figure 13.

[2351] Conclusion:

[2352] There are no visible signs of expression of Coll a as the expected theoretical band size (~150 kDa), however, there is a clear reaction to the His-tag at around 100 kDa, which corresponds to type I collagen.

[2353] Improvement of transfection in tobacco leaves may lead to better positive results in the expression and identification of collagen.

[2354] Example 5 - Purification of Collagen Collagen including a His-tag was prepared from P. pastoris as laid out above. This allows His-tag purification of Collagen from P. pastoris (20 ml) by means of affinity chromatography. The results are depicted in Figure 1 . As expected, collagen was not visible at 280 nm.

[2355] A pellet from the produced batch of the P. pastoris (previously confirmed collagen expression by His WB) was lysated by sonication and used for His purification. Total volume was 5 mL.

[2356] Gel electrophoretic results are shown in Figures 14 and 15. SDS PAGE showed a band at ca. 130 kDa, and Western Blot using His antibody confirmed positive bands for Pichia pastoris samples, while no band was detected for albumin negative control. Clear Purification of initial Material (see SDS PAGE Lane 3) with the major impurities located in Lane 4+5 (Flowthrough) and Collagen in the Elution part (Lane 6 + 7 + 8). No A280nm signal was detected by the FPLC during elution phase, which made the choice of fractions hard. This is repeated for additional elution fractions (D10, D11 , D12 ... .) to check for presence of collagen.

[2357] The purified collagen from Pichia pastoris was further investigated in an SDS PAGE with a gel of 7.5% Tris-glycine, a sample of 10 pl in 3.3 pl of LDS sample buffer and a Coomassie PageBlue stain. The results are depicted in Figure 16.

[2358] The sample was further investigated for sample / protein identity by mass spectrometry. The results are as follows:

[2359]

[2360] 

[2361] The hydroxylation pattern was investigated. The detected peptides were aligned to the sequence of recombinant collagen. Hydroxylated amino acid residues were detected. Besides proline hydroxylation, there is also a methionine hydroxylation that occurs during the preparation of samples. It was found that methionine hydroxylation is rather frequent. His-tag was not detected. These results are shown in Figure 2.

[2362] The cultivating conditions of Pichia pastoris were optimized. It was found that cultivation at 20°C was better than cultivation at higher temperatures such as 28°C. The Western Blot analysis results (nitrocellulose (NC) membrane, 1000 V, 55 min, blocking with Li-Cor TBS, + 5% milk for 1 hour, detection with His-antibody conjugated with horseradish peroxidase (HRP)) are depicted in Figure 17.

[2363] In a further development of the described purification of collagen, not only P. pastoris itself was used, but purification was also carried out from its supernatant. Thus, POI secreted by P. pastoris, here collagen, can also be obtained, increasing the overall yields. A procedure of the adapted process is depicted in Figure 18B and includes: collecting the supernatant from different batches; treatmeant with AS 40% (1 h), precipitation. The obtained pellet is resuspended in extraction buffer (10-15% of the initial volume).

[2364] The supernatant is treated again with AS 60% (1 h) and precipitated, the resulting pellet resuspended in extraction buffer (10-15% of the initial volume). Possible aggregates are removed from the supernatant and the final supernatant is obtained. All fractions were analyzed using SDS PAGE (Figure 20A) and Western blot (Figure 20B). AS 40% precipitation was able to concentrate most of the proteins and a collagen signal was detected in these AS 40% samples. Even though AS 60% and aggregates showed bands, no signal was clearly detected (Figure 20A). Collagen was analysed under denaturing conditions, resulting in a single Band at around 100 kDa in the western blot (Figure 20B).

[2365] Conclusions:

[2366] Clones A8 and A11 express Collagen (intracellular) as stable clones. Purification with HisTrap column works efficiently, 20°C is the better cultivation temperature. Collagen was not only found intracellularly, but also in the supernatant and can be harvested to further increase yields.

[2367] Example 6 - Comparison of different harvesting and purification methods of collagen from plants Different harvesting and purification methods were compared with each other.

[2368] (i) Method 1 :

[2369] Harvesting: Tobacco leaves frozen in liquid nitrogen, grounded and stored at -80°C.

[2370] Extraction: 100 mM Tris, pH 7.5 for 1 h at 4°C. Sample-buffer ratio 1 :3

[2371] Purification:

[2372] (1 ) AS 40% (1 h) and AS 60% (1 h) precipitation. Pellet resuspended in extraction buffer (10-15% initial volume).

[2373] (2) His purification. Collagen was detected in Flowthrough.

[2374] Result: No collagen was detected when 50 g batch was tested.

[2375] (ii) Method 2:

[2376] Harvesting: Production of recombinant collagens by plants, in particular single-chain collagen type I a1 (I) and their use.

[2377] Extraction: Acid Extraction 0.5 M Acetic Acid

[2378] Purification:

[2379] (1 ) NaCI 0.7 M, NaCI 0.9 M; Pellet resuspended in 0.5 M Acetic Acid.

[2380] (2) Samples insufficiently pure: dialysis in 0.5 M Acetic acid + 2nd.

[2381] (3) In case of further purification step: ion exchange (gradient elution 0 - 0.5 M NaCI).

[2382] Result: After precipitation with 0.7 M NaCI, then with 0.9 M NaCI the homotrimeric 80 to 90% pure collagen is present in the 0.9 M NaCI precipitate.

[2383] (iii) Method 3:

[2384] Harvesting: 1 kg of tobacco leaves grounded with 2 L chilled extraction buffer.

[2385] Extraction: 100 mM sodium phosphate buffer pH 7.5, 4.5 mM potassium Meta disulfite, 12.23 mM L-cystein and 7.5 mM EDTA.

[2386] Purification:

[2387] (1 ) 6.68 g charcoal and 16.67 g of PVPP were added to the extract and continuously stirred for 20 minutes.

[2388] (2) AS 15% (1 h) and AS 25% (1 h). Pellet resuspended in extraction buffer.

[2389] (3) Samples were digested with ficin (3 h) and precipitated with two rounds of 3 M NaCI (O.N.).

[2390] (4) After precipitation, final pellet was resuspended in 10 mM HCI and dialysed against 10 mM HCI. Samples were filtered and concentrated.

[2391] Result: After digestion with ficin, the atelocollagen maintains its ability to form fibrils (at least 70%). (iv) Method 4:

[2392] Harvesting: 450 g of tobacco leaves blended in 900 mL chilled extraction buffer.

[2393] Extraction: 100 mM Tris-HCI, pH 7.5 containing 4.5 mM potassium meta bisulfite and 7.5 mM EDTA.

[2394] Purification:

[2395] (1 ) 3 g charcoal and 7.5 g of PVPP were added to the extract. Blending was performed in five intervals of 1 min each.

[2396] (2) Extract was filtered through gauze pad, centrifuged and 10 mM CaCI + 1 g / L activated carbon was added.

[2397] (3) Samples were digested with ficin and precipitated with 3.13 M NaCI (O.N.). Pellet was resuspended in 0.25 M Acetic acid + 2 M NaCI for precipitation. Final pellet: 0.5 M Acetic acid.

[2398] (4) Sample passed through 12 layers of gauze pad and precipitated with 3 M NaCI. Final pellet was resuspended in 10 mM HCI and dialysed against 10 mM HCI. Samples were filtered and concentrated.

[2399] Result: Human procollagen type I in plants by coexpression of human procollagen alpha 1 and alpha 2 chains together with human enzymes P4H alpha, P4H beta, and LH3.

[2400] Example 7 - Downstream processing exemplified for collagen from Pichia pastoris

[2401] A general downstream processing flow schemes are depicted in Figures 18A and 19.

[2402] Table 4. Exemplified process conditions

[2403] In the general scheme as depicted in Figure 19, Tris (PVPP+EDTA) + NaCI precipitation of 1 g plant material and 1.1 mg collagen Pichia is used.

[2404] An SDS PAGE gel of the process as depicted in Figure 19 is shown in Figure 21 A. Herein, a gel “Mini Protean TGX 7.5% Tris-glycine”, a sample loading buffer of 10 pl sample in 3.3 pl LDS sample buffer and a Coomassie (Page blue) stain was used.

[2405] A Western Blot of the process as depicted in Figure 19 is shown in Figure 21 B. A membrane 7.5% Tris-glycine gel, NC membrane was used. The method iBIot P0 was conducted for 7 min. Ll-Cor TBS + 5% milk was used for blocking for 1 hour. Detection was performed by His antibody with horseradish peroxidase (HRP), 1 :1000.

[2406] After previous large His-tag purification from Pichia pastoris, the peak containing the collagen molecule showed additional bands (impurities). Size Exclusion Chromatography (SEC) and 3.13 M NaCI precipitation were tested to check protein profile separation. The results are depicted in Figures 22 and 23. It was found that SEC was successfully able to separate collagen from other proteins (Fractions B8- B6). Since collagen does not absorb at 280 nm, A6-B9 fractions were also tested but no band was detected in SDS PAGE. Protein quantification (A205 nm) and comparison to precipitation recovery may be conducted additionally. Purified Pichia Collagen was detected after spiking plant material. Gelation can be determined in rheological assays.

[2407] Example 8 - Collagen purification from plants

[2408] A general process is shown in the flow scheme of Figure 24.

[2409] For analysis, different approaches were tested. In a first approach, plant material, more spefically plant leaves were collected, ground in liquid nitrogen and incubated in buffer, before freezing. Collagen extracted from the mixture using i) 100 mM Tris, pH 7.5 (1 :3). Collagen precipitation was done using successively 40 and 60% ammonium sulfate. For analysis SDS PAGE and Western Blot were performed and a band of 130 kDa (V1 construct, full length, including propeptides) was detected. For comparison, see Figures 25 -29.

[2410] In a second approach, plant material was extracted using 500 mM NaCI and collagen was precipitated using successively 400 and 900 mM NaCI. For analysis SDS PAGE and Western Blot were performed and a band of 130 kDa (V1 construct, full length, including propeptides) was detected. For comparison, see Figures 30, 31.

[2411] It was found that 100 mM Tris, pH 7.5 and AS 60% precipitation gave consistent results for collagen detection compared to Acid extraction and NaCI precipitation. Both 1 :5 and 1 :3 ratios showed positive bands on WB for AS Precipitation method. For scale-up, the less water to process, the better. Therefore, 1 :3 ratio can be selected for further experiments. Positive controls were successfully detected when NC membrane was used. Therefore, this membrane can be selected for further experiments. Final protein purity still needs improvement. Chromatography techniques should increase protein purity. As alternative, investigation of different AS precipitation fractionations (10 - 60%) can be also performed.

[2412] General process conditions are at least 5 g plant material, extraction buffer: 1000 mM Tris, pH 7.5 in a dilution ratio of 1 :3, precipitation in 60% ammonium sulfate, pellet (at least 10x), heating of the sample before conducting SDS PAGE.

[2413] Example 9 - Optimization of Pichia pastoris purification method

[2414] A scheme is shown in Figure 32. SDS PAGE (7.5 Tris-glycine, loading 21 pl sample +6-7 pl LDS buffer, 10 min, 70°C, 4-20% SDS PAGE) and Western Blot (0.45 pm nitrocellulose (NC), 1000 V for 65 min, blocking for 1 hour, detection via His-tag antibody 1 :100, goat anti -mouse 1 :10000) are performed. The results are depicted in Figures 33 and 34.

[2415] No bands were visible in the supernatant of the previous SDS-PAGE.

[2416] Expected MW for Collagen without propeptides in P. pastoris

[2417] Without signal peptide 96.3 kDa

[2418] With signal peptide 105.6 kDa

[2419] Old Pichia Western Blots with His-tag antibody have sometimes shown artifact bands in the upper part of the membrane, but they usually looked less specific. Collagen usually appears larger on gels than its real MW.

[2420] Collagen could be efficiently obtained from Pichia Pastoris.

[2421] Example 10 - Optimization of E. coli purification method

[2422] The method can be adapted to a Gibson assembly method: Linearize pET151 by PCR without the FGF2 sequence. Amplify Col1A1 insert from plant plasmid. Dpnl digest of the original pET151 in the PCR mix. Run the Gibson assembly with the plasmid backbone and insert. Transform E. coli Top10. Colony PCR or Sequencing for verification of obtained clones. Plasmid Mini-Prep. Transformation of E. coli BL21 or Shuffle T7.

[2423] Performance conditions are 98°C - 30 sec, 98°C - 5 sec, 61 °C - 5 sec, 72°C - 45 sec, 72°C - 1 min, and 4°C.

[2424] The results of an E. coli Top10 Colony-PCR after Gibson Assembly were investigated in a gel electrophoresis as depicted in Figure 35 with Primers 77 / 78, expected band sizes Col1A1 ca. 3.200 bp.

[2425] The results of SDS PAGE (3-8% Tris-acetate, sample loading 10 pl sample + 3.3 pl criterion SB, staining with Coomassie (Page Blue)) and Western Blot (0.45 pm PVDF membrane, 100 V for 55 min, blocking with Li-Cor TBS + 5% milk filtered for 1 hour, detection via Collagen detection kit) are depicted in Figure 36.

[2426] Collagen could be efficiently obtained from E. coli.

[2427] Example 11 - Downstream processing and upscaling of the processes

[2428] It was considered to improve recombinant polypeptide such as collagen production in P. pastoris on a 100-1000 L scale followed by purification, and downstream processing of plant biomass on 100-1000 kg scale.

[2429] Table 5. P. pastoris in comparison

[2430] Basic characteristics of different host systems for the expression of recombinant proteins

[2431] Characteristics Escherichia coli Pichia pastoris CHO cell

[2432] Doubling time 30 min 60-120 min 24 hr

[2433] Cost of growth medium Low Low High

[2434] Complexity of growth Minimum Minimum Complex medium

[2435] Expression level High Low to high Low to moderate

[2436] Extracellular expression Secretion to periplasm Secretion to medium Secretion to medium

[2437] Protein folding Refolding usually required Refolding may be required Proper folding

[2438] N-linked glycosylation None High mannose Complex

[2439] O-linked glycosylation No Yes Yes

[2440] Phosphorylation & No Yes Yes acetylation

[2441] Drawback Accumulation of LPS Codon bias Contamination with animal viruses

[2442] Abbreviations: CHO, Chinese hamster ovary; LPS, lipopolysaccharide.

[2443] Reference: Mohsen Karbalaei, Seyed A. Rezaee, and Hadi Farsiani Pichia pastoris: A highly successful expression system for optima synthesis of heterologous proteins. 2020 Sep; 235(9): 5867-5881. Link: ohttps: / / www.ncbi. nlm.nih.gov / pmc / articles / PMC7228273 /

[2444] Table 6. SWOT analysis of P. pastoris platform technology

[2445] Strengths Weaknesses and Threats Opportunities

[2446] 1. Generally recognized as safe (GRAS * status), robust organism

[2447] 2. Innate ability to secrete heterologous proteins

[2448] 3. A highly inducible promoter (alcohol oxidase) that can be easily exploited for recombinant protein 1 . Low cellular productivity if not optimised production

[2449] 4., No Crabtree effects 2. Protease release during fermentation 1. Improved cellular productivity

[2450] 5. Ability to perform certain post-translational 3. Scale up requires large volume methanol handling ations and high oxygen i 2. Continuous culture with lower cell concentration modific nput together with substantial heat generation and higher productivity, smaller footprint

[2451] 6. Suitable for platform manufacture 4. Harvesting / dewatering of very high cell density results in instability in centrifugation 3. Methanol-free systems

[2452] 7. Very high cell density achievable

[2453] 8. Low Cost of Goods compared to e.g., mammalian systems

[2454] 9. Relatively low secreted host cell protein (HCP)

[2455] 10. Absence of endotoxins / bacterio-phage contamination

[2456] Reference: Salome de Sa Magalhaes andEli Keshavarz-Moore: Pichia pastoris (Komagataella phaffii) as a Cost-Effective Tool fo Vaccine Production for Low- and Middle-Income Countries (LMICs). Bioengineering 2021 , 8(9), 119. Link: https: / / www.mdpi.com / 2306 5354 / 8 / 9 / 119.

[2457] Cultivation and induction of protein expression:

[2458] - First cells a grown in a medium with glucose or glycerol to accumulate biomass, then switch to a medium with methanol;

[2459] -Methanol is added for induction in "pulses" (typically 1 -3% (v / v) per 24 h);

[2460] -Good aeration is crucial during the methanol phase, dissolved oxygen should remain above 20%;

[2461] -Pure 02 can be used in high density fermentations;

[2462] -On a small-scale baffled shake-flasks should be used with high shaking speed.

[2463] The number and type of unit operations may vary according to protein characteristics and final product purity required. Some columns can work directly with filtered supernatants. In case of intracellular protein production, a lysis step has to be added.

[2464] For the proteins that are not secreted, the process would have to be complemented by a high-pressure homogenizer or similar equipment to lyse the cells. A detailed process scheme is depicted in Figure 38.

[2465] For instance 60 g plant or more may be used.

[2466] Scaling up from 10 to 100 liters for P. pastoris works. A flow scheme for preparing collagen from plants is shown in Figure 40.

[2467] - 171 -

[2468] Table 7. Exemplified selection of collagen preparation in Nicotiana benthamiana

[2469] Tuse et al. _

[2470] Parameter Unit (1) (2) Walwyn et al. Nandi et al. Alam et al.

[2471] Industry - Pharmaceutical Biofuel Reagent Pharmaceutical Pharmaceutical

[2472] Molecule - Butyrylcholin- Cellulase enzyme Horseradish Monoclonal antibody Antiviral Protein esterase peroxidase enzyme

[2473] Expression - Transient; Transgenic; Transient; Transient; Transient; viral vector system agroinfiltration inducible agroinfiltration agroinfiltration

[2474] Production kg / year 25 3x10A6 5 300 20

[2475] Expression g / kg FW 0.5 4 0.24 1 0.52 system

[2476] Recovery % 20 - 54 65 70

[2477] Purity % >95 - 250 kU / g >95 >99

[2478] CAPEX $ million 92.4 (U / D 3:7) 11 ,5(U / D 10:0) - 122 (U / D 4:6)

[2479] COGS $ / g 1 ,180 (U / D 3:7) 6.9x10A-3 (U / D 1 ,279 (U / D 2:8) 90-121 (U / D 4:6) 105.80 (U / D 6:4)

[2480] 10:0)

[2481] Abbreviations: BChE, butyrylcholinesterase; CAPEX, capital expenditures; COGS, cost of goods sold; HRP, horseradish peroxidase; mAb, monoclonal antibody; U / D, ratio of upstream to downstream costs

[2482] Reference: Matthew J. McNulty, Yuri Gleba, Daniel Tuse, Simone Hahn-Lbbmann, Anatoli Giritch, Somen Nandi, Karen A. McDonald Techno-economic analysis of a plant-based platform for manufacturing antimicrobial proteins for food safety, iotechnology Progress 2020;36:e2896. https: / / doi.Org / 10.1002 / btpr.2896. Link: https: / / aiche.onlinelibrary.wiley.eom / doi / pdf / 10.1002 / btpr.2896.

[2483] A detailed flow scheme for an example of upscaling is depicted in Figure 41 .

[2484] In gerenal, the upscaled purification process for 1 kg plant mass would encompass the following steps:

[2485] 1 . 1 kg of transgenic tobacco leaves are ground with pre chilled 2 liters extraction buffer in a 4 L reactor (ESCO model EL-3) for 20 minutes;

[2486] 2. Ammonium sulphate precipitation;

[2487] 3. Centrifugation;

[2488] 4. 3 M NaCI precipitation;

[2489] 5. Filtration.

[2490] Harvesting and Collection can be done as described below and published in, e.g., WO 2009 / 053985 A1 and referenced in Fig. 37- 41 .

[2491] Collect a substantial amount of plant leaves from your source, ensuring that they are fresh and healthy.

[2492] Cleaning and Preparation:

[2493] Remove any dirt, debris, or contaminants from the leaves. Wash the leaves with water to eliminate surface impurities.

[2494] Homogenization:

[2495] Grind or homogenize the cleaned leaves to create a plant leaf extract. You can use a large-scale blender, grinder, homogenizer or double stack disintegrator for this purpose.

[2496] Extraction:

[2497] Transfer the homogenized leaf material into a suitable extraction buffer. The choice of buffer may depend on the type of proteins you are targeting.

[2498] Extract the proteins by shaking, stirring, or agitating the mixture. This step helps to solubilize the proteins into the buffer.

[2499] Filtration:

[2500] Exemplified process in detail:

[2501] Filter the plant extract to remove any remaining solid particles or debris, obtaining a clear protein-containing solution.

[2502] (Optional Centrifugation:)

[2503] Centrifuge the protein extract to separate the soluble proteins from cell debris and other insoluble components. The supernatant, containing the soluble proteins, is collected.

[2504] Precipitation or Salting Out: Depending on the proteins of interest, you may need to precipitate them using techniques like ammonium sulfate precipitation or acetone precipitation. This step helps to concentrate the proteins.

[2505] (Optional) Chromatography:

[2506] Perform chromatography, such as size exclusion chromatography, ion exchange chromatography, or affinity chromatography, to further purify and separate the target proteins based on their properties.

[2507] Elution and Collection:

[2508] Elute the purified proteins from the chromatography columns, collecting fractions containing the target proteins.

[2509] Dialysis or Buffer Exchange:

[2510] Dialyze the protein fractions or perform buffer exchange to transfer the purified proteins into a suitable storage or assay buffer while removing unwanted substances.

[2511] Concentration:

[2512] Concentrate the purified protein solutions to achieve the desired concentration, if necessary.

[2513] Analysis and Characterization:

[2514] Analyze the purified proteins for purity, concentration, and integrity using techniques like SDS-PAGE, Western blotting, or mass spectrometry.

[2515] Storage:

[2516] Store the purified proteins in aliquots at appropriate temperatures and conditions to maintain their stability and activity.

[2517] Processing greenhouse-propagated plants provides 25-75 g purified product at an expected yield of 250 mg / kg fresh weight of plant materials.

[2518] Example 12 - Determining the rheological properties of collagen

[2519] It has been verified that collagen has significant thickening properties.

[2520] Method establishment for testing and analysing the gelation of collagen.

[2521] The measurements were conducted using an Anton Paar MCR 90 Rheometer with a CP25-1 (25 mm, 1 °) probe attached. PureCol™ 5 mg / mL was used for a proof-of- concept method establishment to measure the gel strength of a heat induced gelation of collagen. Prior to measurement, the sample needs be cooled at 4°C and a neutral pH 7 + / - 0.5 must be ensured. The measuring plate was set to 4°C before putting 150 pL of the cooled collagen on the surface. The surrounding area was covered with water and a self-made solvent trap (yoghurt cup) was placed around the sample and the probe without touching it. The principle of measurement is depicted in Figure 6.

[2522] Within the method a pre-shear with a shear rate y* = 0.5 [1 / s] was applied to ensure homogeneity. The shear test was conducted using two intervals. The first interval uses a shear strain (oscillating) y = 3 [%], an angular frequency w = 10 [1 / s] and a linear temperature gradient from 4°C to 37°C in 60 s. The second interval uses a shear strain (oscillating) y = 3 [%], an angular frequency w = 10 [1 / s] and a constant temperature of 37°C for 120 min. The storage modulus G’ [Pa] over time was tracked as output function. The experiment was performed as triplicate. The results are shown in Figure 7.

Claims

Claims1. A non-animal organism expressing a collagen that naturally occurs extracellularly in one or more animal species.

2. The non-animal organism of claim 1 , wherein the non-animal organism comprises the genetic material encoding for the collagen permanently, in particular wherein the genetic material is incorporated in the non-animal organism’s genome.

3. The non-animal organism of any one of claims 1 or 2, wherein the non-animal organism is selected from the group consisting of:(A) a plant, in particular a cultivated plant, in particular tobacco or soy;(B) a fungus, preferably a yeast cell, in particular Pichia pastoris', or(C) a bacterium, in particular an Escherichia coli bacterium.

4. The non-animal organism of any one of claims 1 to 3, wherein the collagen has at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% homology (or identity) of an extracellular matrix polypeptide / protein such as collagen, more preferably wherein the extracellular matrix collagen is selected from mammal extracellular matrix collagen and / or bird extracellular matrix collagen.

5. The non-animal organism of any one of claims 1 to 4, wherein the collagen is a protein that is comprised in vegetarian or vegan animal-based comestible nutrient, in particular a vegetarian animal-based comestible nutrient selected from the group consisting of candy, ice cream, a bakery good, a dip, a diary product, aspic, a vegetarian sausage, cream cheese, jam, spread, margarine and / or a sauce.

6. The non-animal organism of any one of claims 1 to 5, wherein the expressed collagen is located intracellularly in the non-animal organism.

7. The non-animal organism of any one of claims 1 to 6, wherein the expressed collagen is located extracellularly in the non-animal organism.

8. The non-animal organism of any one of claims 1 to 7, wherein the expressed collagen contains a tag, in particular a His-tag.

9. A method for preparing a collagen that naturally occurs extracellularly in one or more animal species, comprising the steps of:(i) providing a non-animal organism comprising genetic information encoding for the collagen that naturally occurs extracellularly in one or more animal species;(ii) expressing the collagen in the non-animal organism and optionally subjecting the collagen to one or more posttranslational modifications.

10. The method of claim 9, further comprising one or more of the following: harvesting the non-animal organism that has expressed the collagen; isolating the collagen from the non-animal organism or cell-culture medium in which the non-animal organism is cultivated; preserving the collagen or composition comprising such from spoiling; and / or freeze-drying or drying of the collagen or composition comprising such.

11. The method of any one of claims 9 or 10, wherein the method comprises purification of the collagen by affinity chromatography.

12. The method of any one of claims 9 to 11 , wherein the method further comprises:(iii) preparing a solution of the collagen in dissolved form, optionally filtering and / or centrifugation of the solution to remove any remaining solid particles or debris and obtaining a clear collagen-containing solution, and optionally precipitating the collagen;(iv) isolating the collagen of step (iii) via chromatography and collecting collagen-containing fractions;(v) optionally subjecting the collagen of step (iv) to dialysis or buffer exchange via a size exclusion chromatography column.

13. The method of any one of claims 9 to 12, wherein the method further comprises isolating the collagen by affinity chromatography, preferably by anantibody and / or by binding to a tag attached to the collagen, in particular a His-tag.

14. A method for preparing a comestible nutrient product, in particular a vegetarian comestible nutrient product, comprising the step of adding the collagen that naturally occurs extracellularly in one or more animal species obtained from a method of any one of claims 9 to 13 to one or more further comestible nutrient ingredients.

15. A comestible nutrient product, in particular a vegetarian comestible nutrient product, comprising at least one collagen that naturally occurs extracellularly in one or more animal species obtained from a method of any one of claims 9