Non-LTR reverse transcription transposon system and application thereof

Abstract

A non-LTR reverse transcription transposition subsystem is provided. Provided are an isolated functional protein encoded by a reverse transcription transposon, a nucleic acid encoding the functional protein, a nucleic acid group, a nucleic acid construct, a composition, a recombinant vector, a recombinant host cell and a kit. The invention further provides a method for introducing the exogenous nucleic acid fragment into the genome of the host cell, a method for editing the genome of the host cell and a method for obtaining the host cell with the genome containing the exogenous nucleic acid fragment. The invention also provides application of the functional protein, the nucleic acid, the nucleic acid group, the nucleic acid construct, the composition, the recombinant vector or the recombinant host cell coded by the reverse transcription transposon to introduction of the exogenous nucleic acid fragment gene into a host cell genome or preparation of a medicine or a preparation.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to Chinese Patent Application No. 202310380978.4 filed with the State Intellectual Property Office of China on April 11, 2023, and the entire contents of this Chinese patent application are hereby incorporated by reference in their entirety for all purposes. Technical Field

[0003] The present application relates to the field of molecular biology, and specifically to a non-LTR retrotransposon system and its use. The present application also specifically relates to: a separated functional protein encoded by a retrotransposon, a nucleic acid encoding the functional protein, a nucleic acid group and a nucleic acid construct, and a composition, a recombinant vector, a recombinant host cell and a kit comprising the functional protein. The present application also specifically relates to: a method for introducing an exogenous nucleic acid fragment into a host cell genome, a method for editing a host cell genome, and a method for obtaining a host cell containing an exogenous nucleic acid fragment in the genome. The present application also specifically relates to the use of the functional protein, the nucleic acid, the nucleic acid group, the nucleic acid construct, the composition, the recombinant vector, or the recombinant host cell in introducing an exogenous nucleic acid fragment gene into the host cell genome, or in the preparation of a drug or preparation for gene therapy, cell therapy, genome research, or stem cell induction and post-induction differentiation. Background Art

[0004] A transposon is a DNA sequence that can be inserted or excised within a genome, thereby transferring its own sequence or a complete copy of its own sequence within or between genomes (Mobile DNA III). Transposons are mainly divided into two categories. Among them, type I retrotransposons use RNA as an intermediate. In the process of moving from a donor site to a new insertion site, type I retrotransposons require the host's RNA polymerase to transcribe the transposable element into RNA, and then the reverse transcriptase domain encoded by the retrotransposon reverse transcribes the RNA copy of the transposable element into DNA, which is inserted into the new site. During the retrotransposition process of this type of retrotransposon, the sequence of the donor site will not be cut from the original site, so multiple copies of the retrotransposable element can exist in the genome.

[0005] According to different replication and integration mechanisms, retrotransposons can be divided into two categories: LTR (long terminal repeat) and non-LTR. The latter is a class of mobile genetic elements widely distributed in the genome of eukaryotic cells. According to the characteristics of the contained nuclease functional domains, non-LTR retrotransposons can be further divided into two categories: apurinic / apyrimidinic nucleases (APE type) and restriction enzyme-like nucleases (RLE type). Among them, the R2 family retrotransposons are the main representatives of RLE type non-LTR retrotransposons. Some R2 retrotransposons have high target site specificity and will specifically integrate into the gene encoding 28srRNA.

[0006] The insertion and integration of large gene fragments have important application value in gene therapy, animal and plant molecular breeding, industrial microbial transformation and other fields. At present, there is a lack of effective tools and systems for the insertion and integration of large gene fragments in industry. In recent years, the scientific community has developed some tools and methods that can insert and integrate large gene fragments, but these methods still have some problems. For example, lentivirus or retrovirus are most commonly used to integrate gene sequences in cell immunotherapy and gene therapy for genetic diseases. Several therapeutic products based on this have been used to treat tumors and genetic disorders (Aiuti, A., Roncarolo, MG and Naldini, L. (2017) Gene therapy for ADA-SCID, the first marketing approval of an ex vivo gene therapy in Europe: paving the road for the next generation of advanced therapy medicinal products. EMBO Mol. Med. 9, 737–740; Aiuti, A. et al. (2009) Gene therapy for immunodeficiency due to adenosine deaminase deficiency. N. Engl. J. Med. 360, 447–458). However, there are some potential limitations in the application of using viruses for large-fragment gene integration: first, the randomness of viral integration in the genome will cause carcinogenic risks; second, the size of the exogenous genes that the virus can carry is also limited, which is not conducive to the transfer of large-fragment therapeutic genes; third, the immunogenicity of the virus may affect the long-term expression of exogenous therapeutic genes and re-administration; fourth, the production of viruses requires the help of living cells, which makes the quality control and downstream processing of such products more complicated and more expensive, and has certain disadvantages in industrialization. Therefore, non-viral large-fragment integration can avoid the various disadvantages brought about by viral integration and become a valuable tool in gene therapy.

[0007] As a non-viral gene integration tool, retrotransposons can achieve integration in the host genome and stable expression of large fragments of exogenous genes through RNA delivery, which can not only reduce negative effects such as immunogenicity, but also can be combined with LNP and other delivery technologies to directly treat target cells in vivo. Due to the lack of relevant retrotransposon tools, retrotransposon technology is rarely used in gene therapy or other fields. Few companies in the world are mining a large number of active retrotransposons and trying to develop them into therapeutic tools. Therefore, it is necessary to quickly discover and obtain more active retrotransposon products, and verify and test their functions to provide more options for the development of gene therapy strategies.

[0008] It should be noted that the methods described in this section are not necessarily methods that have been previously conceived or adopted. Unless otherwise indicated, it should not be assumed that any method described in this section is considered to be prior art simply because it is included in this section. Similarly, unless otherwise indicated, the issues mentioned in this section should not be considered to have been recognized in any prior art. Summary of the invention

[0009] Based on this, in order to seek more advanced and more effective non-viral gene integration tools, the present application provides a non-LTR retrotransposon system and its use. The present application also specifically provides a separated functional protein encoded by a retrotransposon, wherein the functional protein has a functional protein sequence selected from the following (i) or a variant sequence of the aforementioned functional protein having functional protein activity in (ii)-(iv): (i) at least one of the amino acid sequences shown in any one of SEQ ID NOs: 1-94; (ii) at least one of the sequences obtained by deleting, replacing, inserting, or mutating 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids in the amino acid sequence shown in any one of SEQ ID NOs: 1-94; (iii) at least one of the amino acid sequences having at least 70%, 80%, 90%, 95%, or 99% identity with the amino acid sequence shown in any one of SEQ ID NOs: 1-94; and (iv) at least one of the sequences obtained by further fusing the amino acid sequence shown in any one of SEQ ID NOs: 1-94 with other sequences. The functional protein encoded by the retrotransposon provided in the present application has high retrotransposition activity and can provide more options for the development of gene integration tools.

[0010] According to an embodiment of the present application, a separated functional protein encoded by a retrotransposon may be provided, wherein the functional protein comprises an amino acid sequence represented by formula (I):

[0011] C(X 1 ) a C(X 2 )b H(X 3 ) c H(I),

[0012] Wherein, a, b, c are the number of amino acids; C is cysteine; H is histidine; (X 1 ) is any amino acid, and a is 1, 2, 3 or 4; (X 2 ) is any amino acid, and b is 11, 12, 13, 14, 15, 16 or 17; and (X 3 ) is any amino acid, and c is 4.

[0013] According to an embodiment of the present application, a separated functional protein encoded by a retrotransposon may be provided, wherein the functional protein comprises an amino acid sequence represented by formula (II):

[0014] G(X 4 ) d QGD(X 5 ) e S(X 6 ) f F(X 7 ) g D(II),

[0015] Wherein, d, e, f, g are the number of amino acids; D is aspartic acid; F is phenylalanine; G is glycine; Q is glutamine; S is serine; (X 4 ) is any amino acid, and d is 2; (X 5 ) is any amino acid, and e is 2; (X 6 ) is any amino acid, and f is 3; and (X 7 ) is any amino acid, and g is 30, 31, 32, 33, 34, 35 or 36.

[0016] According to an embodiment of the present application, a separated functional protein may be provided, wherein the functional protein comprises an amino acid sequence represented by formula (III):

[0017] C(X 8 ) h C(X 9 ) i E(X 10 ) j H(X 11 ) k C(X 12 ) l RH(X 13 ) m PD(X 14 ) n (X15 )(X 16 ) o K(X 17 ) p Y(III),

[0018] Wherein, h, i, j, k, l, m, n, o, p are the number of amino acids; C is cysteine; D is aspartic acid; E is glutamic acid; H is histidine; K is lysine; R is arginine; P is proline; Y is tyrosine; (X 8 ) is any amino acid, and h is 2, 3 or 4; (X 9 ) is any amino acid, and i is 3, 4, 5, 6, 7, 8, 9 or 10; (X 10 ) is any amino acid, and j is 3; (X 11 ) is any amino acid, and k is 4; (X 12 ) is any amino acid, and l is 9; (X 13 ) is any amino acid, and m is 31, 32, 33, 34, 35 or 36; (X 14 ) is any amino acid, and n is 11, 12, 13, 14, 15, 16, 17, 18 or 19; (X 15 ) is aspartic acid or glutamic acid; (X 16 ) is any amino acid, and o is 15, 16, 17, 18 or 19; and (X 17 ) is any amino acid, and p is 3.

[0019] According to an embodiment of the present application, a separated functional protein encoded by a retrotransposon is provided, wherein the functional protein comprises at least two of the amino acid sequences shown in formula (I), formula (II) and formula (III).

[0020] According to an embodiment of the present application, a separated functional protein encoded by a retrotransposon is provided, wherein the functional protein comprises an amino acid sequence shown in formula (I), formula (II) and formula (III).

[0021] According to an embodiment of the present application, a nucleic acid may be provided, wherein the nucleic acid encodes the functional protein described in the present application.

[0022] According to an embodiment of the present application, a nucleic acid set may be provided, the nucleic acid set comprising a 5' untranslated region, wherein the 5' untranslated region comprises at least one of the nucleotide sequences shown in SEQ ID NOs: 95-188.

[0023] According to an embodiment of the present application, a nucleic acid set may be provided, the nucleic acid set comprising a 3' untranslated region, wherein the 3' untranslated region comprises at least one of the nucleotide sequences shown in SEQ ID NOs: 189-282.

[0024] According to an embodiment of the present application, a nucleic acid group can be provided, which comprises a 5' untranslated region and a 3' untranslated region, wherein the 5' untranslated region comprises a nucleotide sequence shown in any one of SEQ ID NOs: 95-188 or a variant thereof, and the 3' untranslated region comprises a nucleotide sequence shown in any one of SEQ ID NOs: 189-282 or a variant thereof, and the RNA transcribed from the nucleic acid group is capable of binding to a functional protein encoded by a specific retrotransposon.

[0025] According to an embodiment of the present application, a nucleic acid construct may be provided, the nucleic acid construct comprising the nucleic acid described in the present application and / or the nucleic acid group described in the present application.

[0026] According to an embodiment of the present application, a composition may be provided, wherein the composition comprises: a functional protein or a functional fragment thereof encoded by an R2 family retrotransposon, or a nucleic acid encoding the functional protein or a functional fragment thereof, wherein the functional protein or the functional fragment thereof has the function of catalyzing the insertion of an exogenous nucleic acid fragment into a cell genome; and a nucleic acid group, which may be recognized by a functional protein or a functional fragment thereof encoded by a specific retrotransposon.

[0027] According to an embodiment of the present application, a recombinant vector may be provided, wherein the recombinant vector comprises a nucleic acid encoding a functional protein described in the present application, a nucleic acid described in the present application, a nucleic acid group described in the present application, a nucleic acid construct described in the present application, or a composition described in the present application.

[0028] According to an embodiment of the present application, a recombinant host cell can be provided, wherein the recombinant host cell comprises the functional protein described in the present application, the nucleic acid encoding the functional protein described in the present application, the nucleic acid described in the present application, the nucleic acid group described in the present application, the nucleic acid construct described in the present application, the composition described in the present application, or the recombinant vector described in the present application.

[0029] According to an embodiment of the present application, a method for introducing an exogenous nucleic acid fragment into a host cell genome can be provided, wherein the method comprises: delivering the functional protein described in the present application, the nucleic acid encoding the functional protein described in the present application, the nucleic acid described in the present application, the nucleic acid group described in the present application, the nucleic acid construct described in the present application, the composition described in the present application, or the recombinant vector described in the present application into the host cell.

[0030] According to an embodiment of the present application, a method for editing the genome of a host cell may be provided, wherein the method comprises: delivering the functional protein described in the present application, the nucleic acid encoding the functional protein described in the present application, the nucleic acid described in the present application, the nucleic acid group described in the present application, the nucleic acid construct described in the present application, the composition described in the present application, or the recombinant vector described in the present application into the host cell.

[0031] According to an embodiment of the present application, a method for obtaining a host cell whose genome contains an exogenous nucleic acid fragment can be provided, wherein the method comprises: delivering the functional protein described in the present application, the nucleic acid encoding the functional protein described in the present application, the nucleic acid described in the present application, the nucleic acid group described in the present application, the nucleic acid construct described in the present application, the composition described in the present application, or the recombinant vector described in the present application into the host cell.

[0032] According to the embodiments of the present application, the functional protein described in the present application, the nucleic acid encoding the functional protein described in the present application, the nucleic acid described in the present application, the nucleic acid construct described in the present application, the nucleic acid construct described in the present application, the composition described in the present application, the recombinant vector described in the present application, or the recombinant host cell described in the present application for use in introducing exogenous nucleic acid fragments into the host cell genome can be provided.

[0033] According to the embodiments of the present application, the functional protein described in the present application, the nucleic acid encoding the functional protein described in the present application, the nucleic acid described in the present application, the nucleic acid group described in the present application, the nucleic acid construct described in the present application, the composition described in the present application, the recombinant vector described in the present application, or the recombinant host cell described in the present application can be provided for use in gene therapy, cell therapy, genome research, or stem cell induction and post-induced differentiation.

[0034] According to an embodiment of the present application, a kit can be provided, wherein the kit comprises the functional protein described in the present application, the nucleic acid encoding the functional protein described in the present application, the nucleic acid described in the present application, the nucleic acid group described in the present application, the nucleic acid construct described in the present application, the composition described in the present application, the recombinant vector described in the present application, or the recombinant host cell described in the present application.

[0035] It should be understood that the content described in this section is not intended to identify the key or important features of the examples of the present application, nor is it intended to limit the scope of the present application. Other features of the present application will become easily understood through the following description. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] The accompanying drawings exemplarily illustrate the embodiments and constitute a part of the specification, and together with the text description of the specification, are used to explain the exemplary implementation of the embodiments. The embodiments shown are for illustrative purposes only and do not limit the scope of the claims. In all drawings, the same reference numerals refer to similar but not necessarily identical elements.

[0037] Figure 1 A schematic diagram showing the order of elements of the vector in Example 1.

[0038] Figure 2 The results of detecting the retrotransposition activity by amplifying the 3' junction of the 28s rRNA gene using a retrotransposon are shown. Figure 2 The different lanes in the figure correspond to the different retrotransposons in Example 2 (sample codes 1-100 in Table 1).

[0039] Figure 3 Shown are the results of detecting retrotransposition activity using a single copy gene (36b4) as a reference. Figure 3 The different lanes in the figure correspond to the different retrotransposons in Example 3 (sample numbers 3, 4, 5, 7, 11, 14, 19, 20, 21, 23, 26, 28, 29, 43, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 59, 60, 63, 75, 83, 87 and 98 in Table 1).

[0040] Figure 4 Shows Figure 3 Quantitative analysis of the retrotransposition activity detected in .

[0041] Figure 5 Shown are the results of testing the integrity of the retrotransposon by amplifying the 5' junction of the 28s rRNA gene using different loci within the retrotransposon. Figure 5 The different lanes in the figure correspond to the different retrotransposons in Example 4 (sample numbers 3, 4, 5, 7, 11, 14, 19, 20, 21, 22, 23, 26, 28, 29, 43, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 59, 60, 63, 75, 83, 87 and 98 in Table 1). DETAILED DESCRIPTION

[0042] Unless otherwise specified or contradictory to the context, the terms or expressions used herein should be read in conjunction with the overall content of the present disclosure and as understood by those of ordinary skill in the art. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art.

[0043] The terms "nucleic acid" and "polynucleotide" are used interchangeably herein to refer to a polymeric form of nucleotides of any length, including deoxyribonucleotides, ribonucleotides, combinations thereof, and analogs thereof.

[0044] The terms "polypeptide" and "peptide" are used interchangeably in this application to refer to amino acid polymers of any length. Therefore, polypeptides, oligopeptides, proteins, antibodies and enzymes are all included in the definition of polypeptides.

[0045] As described in this application, a "fragment" of a sequence refers to a portion of the sequence. For example, a fragment of a nucleic acid sequence refers to a portion of the nucleic acid sequence, and a fragment of an amino acid sequence refers to a portion of the amino acid sequence.

[0046] As described in this application, a "variant" of a sequence is a polynucleotide or polypeptide that is different from a reference polynucleotide or polypeptide, respectively, but retains basic properties. A typical variant of a polynucleotide is different from another reference polynucleotide in nucleic acid sequence, and the difference in nucleic acid sequence may or may not change the amino acid sequence of the polypeptide encoded by the reference polynucleotide. A typical variant of a polypeptide is different from another reference polypeptide in amino acid sequence. In general, the difference is limited, so the sequences of the reference polypeptide and the variant are generally very similar and identical in many regions. The variant polypeptide and the reference polypeptide may differ in amino acid sequence by one or more substitutions, additions, or deletions in any combination. The substituted or inserted amino acid residues may or may not be residues encoded by the genetic code. Variants of polynucleotides or polypeptides may occur naturally, such as allelic variations, or may be unknown naturally occurring variants. Non-naturally occurring polynucleotide and polypeptide variants can be made by mutagenesis techniques, direct synthesis, and other recombinant methods known to skilled technicians.

[0047] The expression "functional protein encoded by a retrotransposon" as used in the present application refers to a polypeptide that catalyzes the integration of an exogenous nucleic acid fragment into a target site (such as genomic or extrachromosomal DNA).

[0048] The term "exogenous nucleic acid fragment" as used in this application includes any gene of interest or any gene or fragment thereof that can be transposed. In some non-limiting embodiments, the exogenous nucleic acid fragment is different from the source of the host cell, for example, a nucleic acid sequence isolated from an organism different from the host cell, i.e., an exogenous nucleic acid fragment relative to the host cell.

[0049] The terms "domain" and "functional domain" used in this application are used interchangeably and refer to the structure of a biomolecule that contributes to a specific function of the biomolecule and may include a continuous region (e.g., a continuous sequence) or different non-continuous regions (e.g., a non-continuous sequence) of the biomolecule. Examples of protein functional domains include, but are not limited to, DNA binding domains, RNA binding domains, reverse transcriptase functional domains, and nuclease functional domains.

[0050] The term "untranslated region" used in this application refers to a nucleic acid sequence located at both ends of a transposable element and flanking a transposable nucleic acid sequence. Among them, the untranslated region located at the 5' end of the transposable nucleic acid sequence is called the 5' untranslated region, and the untranslated region located at the 3' end of the transposable nucleic acid sequence is called the 3' untranslated region. In some embodiments, RNA transcribed from the untranslated region can bind to a functional protein encoded by a specific retrotransposon.

[0051] The term "nucleic acid construct" used in the present application is defined herein as a single-stranded or double-stranded nucleic acid molecule, preferably an artificially constructed nucleic acid molecule. Optionally, the nucleic acid construct also contains one or more regulatory sequences that are operably connected, and the one or more regulatory sequences can guide the coding sequence to be expressed in a suitable host cell under its compatible conditions. The term "expression" should be understood to include any step involved in the production of proteins or polypeptides, including but not limited to transcription, post-transcriptional modification, translation, post-translational modification and secretion. The term "regulatory sequence" includes all components necessary or advantageous for expressing the polypeptide / protein of the present application. Each regulatory sequence can be naturally contained or exogenous to the nucleic acid sequence encoding the protein or polypeptide. These regulatory sequences include but are not limited to leader sequences, polyadenylation sequences, propeptide sequences, promoters, signal sequences and transcription terminators. At a minimum, the regulatory sequence should include a promoter and start and stop signals for transcription and translation. In order to introduce specific restriction sites so that the regulatory sequence can be connected to the coding region of the nucleic acid sequence encoding the protein or polypeptide, a regulatory sequence with a joint can be provided.

[0052] The term "promoter" used in the present application refers to a polynucleotide sequence that can control the transcription of a coding sequence. The promoter sequence includes a specific sequence that is sufficient to enable RNA polymerase recognition, binding and transcription initiation. In addition, the promoter sequence can include a sequence that optionally regulates the recognition, binding and transcription initiation activity of RNA polymerase in the nucleic acid construct provided by the present application. The promoter can affect the transcription of a gene that is located on the same nucleic acid molecule as the promoter or a gene that is located on a different nucleic acid molecule as the promoter.

[0053] The term "host cell" used in this application includes, but is not limited to, animal cells, plant cells, algae cells, fungal cells, yeast cells, or bacterial cells. This term includes the offspring of the original cell into which the exogenous nucleic acid fragment has been introduced. Exemplary host cells include human embryonic kidney cells HEK293T. It should be understood that the offspring of a single parental cell may not be identical to the original parent in morphology or in terms of genome or total DNA complement due to natural, accidental or intentional mutations.

[0054] The term "vector" as used in this application refers to a nucleic acid molecule capable of transporting another nucleic acid molecule to which it is attached. Examples of vectors include, but are not limited to, plasmids, viruses, bacteria, bacteriophages, and insertable DNA fragments. The term "plasmid" refers to a circular double-stranded DNA that can accept exogenous nucleic acid fragments and can replicate in prokaryotic or eukaryotic cells.

[0055] Functional proteins encoded by retrotransposons

[0056] The present application provides a non-LTR retrotransposon system and its use. According to an embodiment of the present application, a separated functional protein encoded by a retrotransposon can be provided, wherein the functional protein has a functional protein sequence selected from the following (i) or a variant sequence of the aforementioned functional protein having functional protein activity in (ii)-(iv): (i) at least one of the amino acid sequences shown in any one of SEQ ID NOs: 1-94; (ii) at least one of the sequences obtained by deleting, replacing, inserting, or mutating 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids in the amino acid sequence shown in any one of SEQ ID NOs: 1-94; (iii) at least one of the amino acid sequences having at least 70%, 80%, 90%, 95%, or 99% identity with the amino acid sequence shown in any one of SEQ ID NOs: 1-94; and (iv) at least one of the sequences obtained by further fusing the amino acid sequence shown in any one of SEQ ID NOs: 1-94 with other sequences.

[0057] According to an embodiment of the present application, a separated functional protein encoded by a retrotransposon may be provided, wherein the functional protein comprises an amino acid sequence represented by formula (I):

[0058] C(X 1 ) a C(X 2 ) b H(X 3 ) c H(I),

[0059] Wherein, a, b, c are the number of amino acids; C is cysteine; H is histidine; (X 1) is any amino acid, and a is 1, 2, 3 or 4; (X 2 ) is any amino acid, and b is 11, 12, 13, 14, 15, 16 or 17; and (X 3 ) is any amino acid, and c is 4.

[0060] According to an embodiment of the present application, a separated functional protein encoded by a retrotransposon may be provided, wherein the functional protein comprises an amino acid sequence represented by formula (II):

[0061] G(X 4 ) d QGD(X 5 ) e S(X 6 ) f F(X 7 ) g D(II),

[0062] Wherein, d, e, f, g are the number of amino acids; D is aspartic acid; F is phenylalanine; G is glycine; Q is glutamine; S is serine; (X 4 ) is any amino acid, and d is 2; (X 5 ) is any amino acid, and e is 2; (X 6 ) is any amino acid, and f is 3; and (X 7 ) is any amino acid, and g is 30, 31, 32, 33, 34, 35 or 36.

[0063] According to an embodiment of the present application, a separated functional protein may be provided, wherein the functional protein comprises an amino acid sequence represented by formula (III):

[0064] C(X 8 ) h C(X 9 ) i E(X 10 ) j H(X 11 ) k C(X 12 ) l RH(X 13 ) m PD(X 14 ) n (X 15 )(X 16 ) o K(X 17 ) p Y(III),

[0065] Wherein, h, i, j, k, l, m, n, o, p are the number of amino acids; C is cysteine; D is aspartic acid; E is glutamic acid; H is histidine; K is lysine; R is arginine; P is proline; Y is tyrosine; (X 8 ) is any amino acid, and h is 2, 3 or 4; (X 9 ) is any amino acid, and i is 3, 4, 5, 6, 7, 8, 9 or 10; (X 10 ) is any amino acid, and j is 3; (X 11 ) is any amino acid, and k is 4; (X 12 ) is any amino acid, and l is 9; (X 13 ) is any amino acid, and m is 31, 32, 33, 34, 35 or 36; (X 14 ) is any amino acid, and n is 11, 12, 13, 14, 15, 16, 17, 18 or 19; (X 15 ) is aspartic acid or glutamic acid; (X 16 ) is any amino acid, and o is 15, 16, 17, 18 or 19; and (X 17 ) is any amino acid, and p is 3.

[0066] According to an embodiment of the present application, a separated functional protein encoded by a retrotransposon is provided, wherein the functional protein comprises at least two of the amino acid sequences shown in formula (I), formula (II) and formula (III).

[0067] According to an embodiment of the present application, a separated functional protein encoded by a retrotransposon is provided, wherein the functional protein comprises an amino acid sequence shown in formula (I), formula (II) and formula (III).

[0068] In some embodiments, the retrotransposon is a non-LTR (non-long terminal repeat) retrotransposon. In some embodiments, the non-LTR retrotransposon is a restriction enzyme-like nuclease (RLE) retrotransposon. In some embodiments, the retrotransposon is an R2 family retrotransposon.

[0069] In some embodiments, the functional protein comprises a DNA binding domain capable of binding to a specific region in the cell genome, preferably, the DNA binding domain is capable of binding to a gene encoding 28s rRNA. In some non-limiting examples, the functional protein uses the 28s rRNA gene as a target site. In some non-limiting examples, the functional protein uses a non-28s rRNA gene as a target site.

[0070] In certain embodiments, the functional protein further comprises an RNA binding domain, a reverse transcriptase functional domain and / or a nuclease functional domain. In some non-limiting examples, the RNA binding domain of the functional protein is combined with the RNA transcribed by the retrotransposon to form an RNP (ribonucleoprotein). In some non-limiting embodiments, the functional protein comprises a nuclease functional domain and / or a reverse transcriptase functional domain. The nuclease functional domain produces a single-stranded gap at the DNA target site, and then utilizes the 3' end of the DNA exposed by the cutting to trigger the reverse transcription of the retrotransposon, wherein the transcribed RNA is used as a template for reverse transcription. Finally, the insertion at the target site is realized by "copy-paste".

[0071] In some embodiments, the species source of the functional protein includes the phylum Arthropoda or Chordata. In some embodiments, the species source of the functional protein includes Accipiter gentilis, Actinemysmarmorata, Agelaius tricolor, Agrochola macilenta, Anagrus nilaparvatae, Andrena haemorrhoa, Anoplius nigerrimus, Artemisiospiza belli, Asobara japonica, Athalia rosae, Blastobasis lacticolella, Bombus hortorum, Bombus hypnorum, Bombus pratorum, Bombus vancouverensis, Brenthis nigerrimus ... ino), Cardiocondylaobscurior, Cerceris rybyensis, Clusia tigrina, Colaptes auratus, Crematogaster levior, Crotalus tigris, Cuora mccordi, Dinocampus coccinellae, Dolichovespula saxonica, Drosophila albomicans, Drosophilasaltans, Ennomos quercinarius, Eopsaltria australis, Eristalis pertinax, Erithacus rubecula, Euphyes dion, Heliconius hecale), Hesperophylax magnus, Ichneumonxanthorius, Jera tricuspidata, Junonia litoralis, Lasioglossumbaleicum), Lasioglossum lativentre, Lasioglossummorio, Leptopilina heterotoma, Lysandra coridon, Marasmarcha lunaedactyla, Mimumesa dahlbomi, Muschampia plurimacula, Neodiprion fabricii, Neodiprion pinetum, Nephrotoma flavescens, Nomada fabriciana, Nylanderia fulva, Nymphalis io, Papilio machaon, Pararge aegeria, Podocnemis expansa, Poecilia wingei, Prinia subflava, Scaptomyza hsui, Schistocerca americana, Seladonia tumulorum, Sphecodes monilicornis, Tibetan loach, Trypoxylus dichotomus, Urbanus tucuti, Venturia canescens, Vespa crabro, Zaprionus camerounensis, or Zaprionus kolodkinae.

[0072] According to an embodiment of the present application, a nucleic acid may be provided, wherein the nucleic acid encodes the functional protein described in the present application.

[0073] Nucleic acid construct

[0074] According to an embodiment of the present application, a nucleic acid set may be provided, the nucleic acid set comprising a 5' untranslated region, wherein the 5' untranslated region comprises at least one of the nucleotide sequences shown in SEQ ID NOs: 95-188.

[0075] According to an embodiment of the present application, a nucleic acid set may be provided, the nucleic acid set comprising a 3' untranslated region, wherein the 3' untranslated region comprises at least one of the nucleotide sequences shown in SEQ ID NOs: 189-282.

[0076] According to an embodiment of the present application, a nucleic acid group can be provided, the nucleic acid group comprising a 5' untranslated region and a 3' untranslated region, wherein the 5' untranslated region comprises a nucleotide sequence shown in any one of SEQ ID NOs: 95-188 or a variant thereof, and the 3' untranslated region comprises a nucleotide sequence shown in any one of SEQ ID NOs: 189-282 or a variant thereof, and the RNA transcribed from the nucleic acid group can bind to a functional protein encoded by a specific retrotransposon. In some embodiments, the RNA transcribed from the untranslated region can recognize and bind to an RNA binding domain of a functional protein encoded by a specific retrotransposon.

[0077] According to an embodiment of the present application, a nucleic acid construct can be provided, which comprises a nucleic acid encoding a functional protein described in the present application and / or a nucleic acid group described in the present application. In some embodiments, the nucleic acid construct further comprises an exogenous nucleic acid fragment. In some embodiments, the exogenous nucleic acid fragment is operably inserted into the nucleic acid construct through a multiple cloning insertion site, and the exogenous nucleic acid fragment may have one or more, which may be the same or different. In some embodiments, the exogenous nucleic acid fragment includes any gene of interest or any gene that can be transposed, preferably, the exogenous nucleic acid fragment includes a natural functional protein gene, an artificial chimeric gene, and / or a non-coding RNA gene. In some embodiments, the non-coding RNA gene includes a variety of known RNAs and RNAs of unknown functions such as rRNA, tRNA, small interfering RNA (siRNA), small nuclear RNA (snRNA), small molecule nucleolar RNA (snoRNA), microRNA (miRNA), and / or long non-coding RNA (lncRNA). In some embodiments, the artificial chimeric gene includes a chimeric antigen receptor gene. In some embodiments, the natural functional protein gene includes a fluorescence-based reporter gene, a luciferase gene, and / or an antibiotic resistance gene. In some non-limiting embodiments, the exogenous nucleic acid fragment may also include a primer set for polymerase chain reaction to facilitate screening for retrotransposition activity.

[0078] In some embodiments, the nucleic acid construct also optionally includes a homologous sequence with 100% identity to at least 10 nucleotides in a specific region in the cell genome, preferably, optionally also includes a homologous sequence with 100% identity to at least 10 nucleotides of a gene encoding 28s rRNA. In some non-limiting embodiments, a part of the homologous sequence has identity with the target site to promote the start of the reverse transcription process. In some non-limiting embodiments, the nucleic acid construct includes 5' untranslated region and 3' untranslated region described in the present application, and the homologous sequence is located at the upstream of the 5' untranslated region and / or the downstream of the 3' untranslated region. In some non-limiting embodiments, the homologous sequence is located at the upstream of the 5' untranslated region. In some non-limiting embodiments, the homologous sequence is located at the downstream of the 3' untranslated region. In some non-limiting embodiments, the homologous sequence is located at the upstream of the 5' untranslated region and the downstream of the 3' untranslated region.

[0079] In some embodiments, the nucleic acid construct further comprises a promoter and a poly (A) sequence. The promoter can be any suitable promoter sequence, i.e., a nucleic acid sequence that can be recognized by the host cell expressing the exogenous nucleic acid fragment. The promoter sequence contains a transcriptional regulatory sequence that mediates protein or polypeptide expression. The promoter can be any nucleic acid sequence that has transcriptional activity in the selected host cell, including mutant, truncated and heterozygous promoters, and can be derived from genes encoding extracellular or intracellular proteins or polypeptides that are homologous or heterologous to the host cell. In some embodiments, the promoter includes CMV, EF1a, SV40, PGK, UbC, human beta actin, CAG, TRE, UAS, Ac5, GFAP, polyhedrin promoter, TBG, ALB, ApoEHCR-hAAT, CaMKIIa, GAL1, TEF1, GDS, ADH1, CaMV35S, Ubi, H1, U6, T7, T7lac, Sp6, araBAD, trp, lac, Ptac, or pL. Poly (A) tailing signal sequences well known in the art and various truncated forms of poly (A) tailing signals can be used in the present application.

[0080] In some embodiments, the nucleic acid construct further comprises any transcription termination sequence (i.e., a sequence that can be recognized by a host cell to terminate transcription) to control the expression of the exogenous nucleic acid fragment. Any terminator that can function in a selected host cell can be used in the present application.

[0081] Optionally, the nucleic acid construct may also include a suitable leader sequence (i.e., an mRNA non-translated region that is very important for the translation of the host cell) to control the expression of the exogenous nucleic acid fragment. The leader sequence may be operably linked to the 5' end of the nucleic acid sequence encoding the polypeptide. Any leader sequence that can function in the selected host cell may be used in the present application.

[0082] Optionally, the nucleic acid construct may further comprise a propeptide coding region to control the expression of the exogenous nucleic acid fragment, the propeptide coding region encoding an amino acid sequence located at the amino terminus of the polypeptide. The resulting polypeptide is referred to as a proenzyme or propolypeptide. Propolypeptides are generally inactive and can be converted into mature active polypeptides by cleaving the propeptide from the propolypeptide by catalysis or autocatalysis.

[0083] Optionally, the nucleic acid construct may also include a regulatory sequence that can regulate the expression of the exogenous nucleic acid fragment according to the growth of the host cell. Examples of regulatory sequences are those that can respond to chemical or physical stimuli (including in the presence of regulatory compounds) to open or close systems for gene expression. Other examples of regulatory sequences are those that can amplify genes. In these examples, the exogenous nucleic acid fragment should be operably linked to the regulatory sequence.

[0084] Retrotransposition composition

[0085] According to an embodiment of the present application, a composition can be provided, wherein the composition comprises: a functional protein or a functional fragment thereof encoded by an R2 family retrotransposon, or a nucleic acid encoding the functional protein or a functional fragment thereof, wherein the functional protein or the functional fragment thereof has the function of catalyzing the insertion of an exogenous nucleic acid fragment into a cell genome; and a nucleic acid group, wherein the RNA transcribed from the nucleic acid group can be recognized by the functional protein or the functional fragment thereof encoded by a specific retrotransposon.

[0086] In some embodiments, the composition is selected from at least one of the following groups (1) to (95), any one of the following groups (1) to (94) comprising: a functional protein-related sequence and a nucleic acid group,

[0087] (1) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 1 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 95; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 189;

[0088] (2) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 2 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 96; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 190;

[0089] (3) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 3 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 97; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 191;

[0090] (4) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 4 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 98; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 192;

[0091] (5) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 5 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 99; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 193;

[0092] (6) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 6 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 100; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 194;

[0093] (7) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 7 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 101; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 195;

[0094] (8) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 8 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 102; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 196;

[0095] (9) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 9 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 103; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 197;

[0096] (10) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 10 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 104; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 198;

[0097] (11) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 11 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 105; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 199;

[0098] (12) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 12 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 106; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 200;

[0099] (13) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 13 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 107; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 201;

[0100] (14) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 14 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 108; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 202;

[0101] (15) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 15 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 109; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 203;

[0102] (16) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 16 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 110; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 204;

[0103] (17) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 17 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 111; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 205;

[0104] (18) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 18 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 112; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 206;

[0105] (19) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 19 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 113; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 207;

[0106] (20) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 20 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 114; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 208;

[0107] (21) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 21 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 115; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 209;

[0108] (22) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 22 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 116; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 210;

[0109] (23) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 23 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 117; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 211;

[0110] (24) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 24 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 118; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 212;

[0111] (25) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 25 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 119; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 213;

[0112] (26) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 26 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 120; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 214;

[0113] (27) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 27 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 121; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 215;

[0114] (28) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 28 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 122; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 216;

[0115] (29) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 29 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 123; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 217;

[0116] (30) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 30 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 124; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 218;

[0117] (31) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 31 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 125; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 219;

[0118] (32) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 32 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 126; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 220;

[0119] (33) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 33 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 127; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 221;

[0120] (34) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 34 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 128; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 222;

[0121] (35) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 35 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 129; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 223;

[0122] (36) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 36 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 130; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 224;

[0123] (37) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 37 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 131; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 225;

[0124] (38) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 38 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 132; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 226;

[0125] (39) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 39 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 133; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 227;

[0126] (40) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 40 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 134; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 228;

[0127] (41) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 41 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 135; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 229;

[0128] (42) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 42 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 136; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 230;

[0129] (43) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 43 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 137; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 231;

[0130] (44) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 44 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 138; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 232;

[0131] (45) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 45 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 139; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 233;

[0132] (46) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 46 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 140; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 234;

[0133] (47) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 47 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 141; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 235;

[0134] (48) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 48 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 142; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 236;

[0135] (49) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 49 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 143; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 237;

[0136] (50) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 50 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 144; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 238;

[0137] (51) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 51 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 145; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 239;

[0138] (52) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 52 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 146; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 240;

[0139] (53) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 53 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 147; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 241;

[0140] (54) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 54 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 148; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 242;

[0141] (55) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 55 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 149; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 243;

[0142] (56) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 56 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 150; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 244;

[0143] (57) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 57 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 151; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 245;

[0144] (58) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 58 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 152; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 246;

[0145] (59) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 59 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 153; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 247;

[0146] (60) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 60 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 154; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 248;

[0147] (61) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 61 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 155; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 249;

[0148] (62) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 62 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 156; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 250;

[0149] (63) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 63 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 157; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 251;

[0150] (64) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 64 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 158; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 252;

[0151] (65) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 65 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 159; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 253;

[0152] (66) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 66 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 160; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 254;

[0153] (67) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 67 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 161; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 255;

[0154] (68) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 68 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 162; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 256;

[0155] (69) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 69 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 163; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 257;

[0156] (70) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 70 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 164; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 258;

[0157] (71) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 71 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 165; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 259;

[0158] (72) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 72 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 166; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 260;

[0159] (73) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 73 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 167; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 261;

[0160] (74) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 74 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 168; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 262;

[0161] (75) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 75 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 169; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 263;

[0162] (76) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 76 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 170; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 264;

[0163] (77) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 77 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 171; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 265;

[0164] (78) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 78 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 172; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 266;

[0165] (79) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 79 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 173; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 267;

[0166] (80) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 80 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 174; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 268;

[0167] (81) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 81 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 175; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 269;

[0168] (82) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 82 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 176; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 270;

[0169] (83) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 83 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 177; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 271;

[0170] (84) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 84 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 178; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 272;

[0171] (85) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 85 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 179; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 273;

[0172] (86) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 86 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 180; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 274;

[0173] (87) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 87 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 181; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 275;

[0174] (88) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 88 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 182; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 276;

[0175] (89) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 89 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 183; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 277;

[0176] (90) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 90 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 184; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 278;

[0177] (91) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 91 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 185; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 279;

[0178] (92) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 92 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 186; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 280;

[0179] (93) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 93 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 187; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 281;

[0180] (94) the functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 94 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 188; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 282; or

[0181] (95) A variant of any one of the aforementioned groups (1) to (94),

[0182] Among them, the functional protein-related sequence is the amino acid sequence of the variant of each group of functional proteins or the nucleic acid sequence encoding the variant, and the variant has a variant sequence of the aforementioned functional protein with functional protein activity selected from the following (i)-(iii): (i) at least one of the sequences obtained by deleting, replacing, inserting, or mutating 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids in the amino acid sequence of each group of functional proteins; (ii) at least one of the amino acid sequences that have at least 70%, 80%, 90%, 95% or 99% identity with the amino acid sequence shown in any one of SEQ ID NOs: 1-94; and (iii) at least one of the sequences obtained by further fusing other sequences with the amino acid sequence shown in any one of SEQ ID NOs: 1-94.

[0183] In some embodiments, the nucleic acid group also includes an exogenous nucleic acid fragment. In some embodiments, the exogenous nucleic acid fragment is operably inserted into the nucleic acid construct through a multiple cloning insertion site, and the exogenous nucleic acid fragment may have one or more, which may be the same or different. In some embodiments, the exogenous nucleic acid fragment includes any gene of interest or any gene that can be transposed, preferably, the exogenous nucleic acid fragment includes a natural functional protein gene, an artificial chimeric gene, and / or a non-coding RNA gene. In some embodiments, non-coding RNA genes include RNAs of various known functions such as rRNA, tRNA, small interfering RNA (siRNA), small nuclear RNA (snRNA), small molecule nucleolar RNA (snoRNA), microRNA (miRNA), and / or long non-coding RNA (lncRNA) and RNAs of unknown functions. In some embodiments, artificial chimeric genes include chimeric antigen receptor genes. In some embodiments, natural functional protein genes include fluorescence-based reporter genes, luciferase genes, and / or resistance genes. In some non-limiting embodiments, the exogenous nucleic acid fragment may also include a primer series for polymerase chain reaction to facilitate screening for reverse transcription transposition activity.

[0184] In some embodiments, the nucleic acid group also optionally includes a homologous sequence with 100% identity to at least 10 nucleotides in a specific region in the cell genome, preferably, optionally also includes a homologous sequence with 100% identity to at least 10 nucleotides of a gene encoding 28srRNA. In some non-limiting embodiments, a part of the homologous sequence has identity with the target site to promote the start of the reverse transcription process. In some non-limiting embodiments, the nucleic acid construct includes 5' untranslated region and 3' untranslated region described in the present application, and the homologous sequence is located upstream of the 5' untranslated region and / or downstream of the 3' untranslated region. In some non-limiting embodiments, the homologous sequence is located upstream of the 5' untranslated region. In some non-limiting embodiments, the homologous sequence is located downstream of the 3' untranslated region. In some non-limiting embodiments, the homologous sequence is located upstream of the 5' untranslated region and downstream of the 3' untranslated region.

[0185] In certain embodiments, nucleic acid and / or nucleic acid group also comprise promoter and poly (A) sequence.Promoter can be any suitable promoter sequence, i.e., a nucleic acid sequence that can be recognized by the host cell expressing nucleic acid sequence.Promoter sequence contains transcriptional regulatory sequence that mediates protein or polypeptide expression.Promoter can be any nucleic acid sequence with transcriptional activity in selected host cell, including mutant, truncated and heterozygous promoter, which can be derived from genes encoding extracellular or intracellular proteins or polypeptides homologous or heterologous to host cells.In certain embodiments, promoter includes CMV, EF1a, SV40, PGK, UbC, human beta actin, CAG, TRE, UAS, Ac5, GFAP, polyhedrin promoter, TBG, ALB, ApoEHCR-hAAT, CaMKIIa, GAL1, TEF1, GDS, ADH1, CaMV35S, Ubi, H1, U6, T7, T7lac, Sp6, araBAD, trp, lac, Ptac or pL. Poly (A) tailing signal sequences well known in the art and various truncated forms of poly (A) tailing signals can be used in the present application.

[0186] In some embodiments, the nucleic acid and / or nucleic acid group encoding the amino acid sequence also includes any transcription termination sequence, i.e., a sequence that can be recognized by the host cell to terminate transcription. The termination sequence is operably connected to the 3' end of the nucleic acid sequence encoding the protein or polypeptide. Any terminator that can function in the selected host cell can be used in the present application.

[0187] Optionally, the nucleic acid and / or nucleic acid group encoding the amino acid sequence may also include a suitable leader sequence, i.e., an mRNA non-translational region that is very important for translation of the host cell. The leader sequence may be operably linked to the 5' end of the nucleic acid sequence encoding the polypeptide. Any leader sequence that may function in the selected host cell may be used in the present application.

[0188] Optionally, the nucleic acid and / or nucleic acid group encoding the amino acid sequence may also include a propeptide coding region, which encodes the amino acid sequence located at the amino terminus of the polypeptide. The resulting polypeptide is referred to as a proenzyme or propolypeptide. Propolypeptides are generally inactive and can be converted into mature active polypeptides by cleaving the propeptide from the propolypeptide by catalysis or autocatalysis.

[0189] Optionally, the nucleic acid and / or nucleic acid group encoding the amino acid sequence may also include a regulatory sequence that can regulate the expression of the polypeptide according to the growth of the host cell. Examples of regulatory sequences are those that can respond to chemical or physical stimuli (including in the presence of regulatory compounds) to open or close systems for gene expression. Other examples of regulatory sequences are those that can amplify genes. In these examples, the nucleic acid sequence encoding the protein or polypeptide should be operably linked to the regulatory sequence.

[0190] Recombinant vectors, recombinant host cells and kits

[0191] According to an embodiment of the present application, a recombinant vector may be provided, wherein the recombinant vector comprises a nucleic acid encoding a functional protein described in the present application, a nucleic acid described in the present application, a nucleic acid group described in the present application, a nucleic acid construct described in the present application, or a composition described in the present application. The recombinant vector may be any suitable vector. In some embodiments, the recombinant vector includes, but is not limited to, a recombinant cloning vector, a recombinant eukaryotic expression plasmid, or a recombinant viral vector. In some embodiments, the recombinant eukaryotic expression plasmid includes pcDNA3.1, pCMV, pUC18, pUC19, pUC57, pBAD, pET, pENTR, pGenlenti, or pAAV. In some embodiments, the recombinant viral vector includes a recombinant adenovirus vector, a recombinant adeno-associated virus vector, a recombinant retrovirus vector, a recombinant herpes simplex virus vector, or a recombinant vaccinia virus vector. The recombinant vector of the present application may be constructed by methods well known in the art. For example, appropriate restriction sites may be added to both ends of the nucleic acid construct of the present application, based on the restriction sites contained in the backbone vector used, and then loaded into the backbone vector.

[0192] According to an embodiment of the present application, a recombinant host cell can be provided, wherein the recombinant host cell comprises a functional protein described in the present application, a nucleic acid encoding a functional protein described in the present application, a nucleic acid described in the present application, a nucleic acid group described in the present application, a nucleic acid construct described in the present application, a composition described in the present application, or a recombinant vector described in the present application. The recombinant host cell can be any host cell to which a retrotransposon can be applied. In some embodiments, the recombinant host cell includes but is not limited to an animal cell, a plant cell, an algae cell, a fungal cell, a yeast cell, or a bacterial cell. In some embodiments, the animal cell includes a mammalian cell. In some embodiments, mammalian cells include primary cells (e.g., mesenchymal stem cells, endothelial cells, epithelial cells, fibroblasts, keratinocytes, melanocytes, smooth muscle cells, and immune cells), immortalized cell lines (e.g., HEK293, NIH-3T3, RAW-264.7, STO, VERO, CT26, hTERT immortalized human endothelial / epithelial / fibroblast / keratinocyte / ductal / cell lines), cancer cell lines (e.g., Hela, HepG2 / 3, HL-60, HT-1080, HT-29, A549, SW 620, HCT-15, HCT116, MDA-MB-231, MCF7, SK-OV-3, PANC-1, AsPc-1, THP-1, Huh7, KG-1, RAJI, HB-CB, Jurkat, K562, CRL5826, CHO, MDCK, Renca), embryonic stem cell lines (e.g., H1, H9, WIBR2, WIBR3, G-Olig2, ESF158, RW.4, R1, D3) and differentiated cells thereof, or induced pluripotent stem cell lines and differentiated cells thereof.

[0193] According to an embodiment of the present application, a kit can be provided, wherein the kit comprises the functional protein described in the present application, the nucleic acid encoding the functional protein described in the present application, the nucleic acid described in the present application, the nucleic acid group described in the present application, the nucleic acid construct described in the present application, the composition described in the present application, the recombinant vector described in the present application, or the recombinant host cell described in the present application.

[0194] Methods and uses

[0195] The retrotransposon-based tools and methods for large-fragment gene insertion and integration provided in this application can be applied to multiple fields such as gene therapy, crop breeding, model animal transformation, and industrial microbial transformation. In particular, in the field of gene therapy and cell therapy, these tools and methods can be applied to gene writing therapy, which is of great significance for the treatment of genetic diseases that require long-fragment gene correction.

[0196] According to an embodiment of the present application, a method for introducing an exogenous nucleic acid fragment into a host cell genome can be provided, wherein the method comprises: delivering the functional protein described in the present application, the nucleic acid encoding the functional protein described in the present application, the nucleic acid described in the present application, the nucleic acid group described in the present application, the nucleic acid construct described in the present application, the composition described in the present application, or the recombinant vector described in the present application into the host cell.

[0197] According to an embodiment of the present application, a method for editing the genome of a host cell may be provided, wherein the method comprises: delivering the functional protein described in the present application, the nucleic acid encoding the functional protein described in the present application, the nucleic acid described in the present application, the nucleic acid group described in the present application, the nucleic acid construct described in the present application, the composition described in the present application, or the recombinant vector described in the present application into the host cell.

[0198] According to an embodiment of the present application, a method for obtaining a host cell whose genome contains an exogenous nucleic acid fragment can be provided, wherein the method comprises: delivering the functional protein described in the present application, the nucleic acid encoding the functional protein described in the present application, the nucleic acid described in the present application, the nucleic acid group described in the present application, the nucleic acid construct described in the present application, the composition described in the present application, or the recombinant vector described in the present application into the host cell.

[0199] The method of delivery into the host cell can be any suitable method. In some embodiments, the delivery method includes but is not limited to cationic liposome delivery, lipid nanoparticle delivery, cationic polymer delivery, vesicle-exosome delivery, gold nanoparticle delivery, polypeptide and protein delivery, retrovirus delivery, lentivirus delivery, adenovirus delivery, adeno-associated virus delivery, electroporation delivery, Agrobacterium infection delivery, or gene gun delivery. The cell transfection and culture methods are conventional methods in the art, and suitable transfection and culture methods can be selected according to different cell types.

[0200] The host cell can be any host cell to which the retrotransposon can be applied. In some embodiments, the host cell includes, but is not limited to, an animal cell, a plant cell, an algae cell, a fungal cell, a yeast cell, or a bacterial cell. In some embodiments, the host cell includes a mammalian cell. In some embodiments, the host cell includes a primary cell (e.g., a mesenchymal stem cell, an endothelial cell, an epithelial cell, a fibroblast, a keratinocyte, a melanocyte, a smooth muscle cell, an immune cell), an immortalized cell line (e.g., HEK293, NIH-3T3, RAW-264.7, STO, VERO, CT26, hTERT immortalized human endothelial / epithelial / fibroblast / keratinocyte / ductal / cell line), a cancer cell line (e.g., Hela, HepG2 / 3, HL-60, HT-1080, HT-29, A549, SW62, 0, HCT-15, HCT116, MDA-MB-231, MCF7, SK-OV-3, PANC-1, AsPc-1, THP-1, Huh7, KG-1, RAJI, HB-CB, Jurkat, K562, CRL5826, CHO, MDCK, Renca), embryonic stem cell lines (e.g., H1, H9, WIBR2, WIBR3, G-Olig2, ESF158, RW.4, R1, D3) and differentiated cells thereof, or induced pluripotent stem cell lines and differentiated cells thereof.

[0201] According to the embodiments of the present application, the functional protein described in the present application, the nucleic acid encoding the functional protein described in the present application, the nucleic acid described in the present application, the nucleic acid group described in the present application, the nucleic acid construct described in the present application, the composition described in the present application, the recombinant vector described in the present application, or the recombinant host cell described in the present application can be provided for the purpose of introducing the exogenous nucleic acid fragment into the host cell genome. The host cell can be any host cell that the retrotransposon can be applied. In certain embodiments, the host cell includes but is not limited to animal cells, plant cells, algae cells, fungal cells, yeast cells, or bacterial cells. In certain embodiments, the host cell includes mammalian cells. In some embodiments, host cells include primary cells (e.g., mesenchymal stem cells, endothelial cells, epithelial cells, fibroblasts, keratinocytes, melanocytes, smooth muscle cells, immune cells), immortalized cell lines (e.g., HEK293, NIH-3T3, RAW-264.7, STO, VERO, CT26, hTERT immortalized human endothelial / epithelial / fibroblast / keratinocyte / ductal / cell lines), cancer cell lines (e.g., Hela, HepG2 / 3, HL-60, HT-1080, HT-29, A549, SW62 0, HCT-15, HCT116, MDA-MB-231, MCF7, SK-OV-3, PANC-1, AsPc-1, THP-1, Huh7, KG-1, RAJI, HB-CB, Jurkat, K562, CRL5826, CHO, MDCK, Renca), embryonic stem cell lines (e.g., H1, H9, WIBR2, WIBR3, G-Olig2, ESF158, RW.4, R1, D3) and differentiated cells thereof, or induced pluripotent stem cell lines and differentiated cells thereof.

[0202] According to the embodiments of the present application, the functional protein described in the present application, the nucleic acid encoding the functional protein described in the present application, the nucleic acid described in the present application, the nucleic acid group described in the present application, the nucleic acid construct described in the present application, the composition described in the present application, the recombinant vector described in the present application, or the recombinant host cell described in the present application can be provided for use in gene therapy, cell therapy, genome research, or stem cell induction and post-induced differentiation.

[0203] The various embodiments and preferences for the present application described above may be combined with each other (as long as they are not inherently contradictory to each other and are applicable to the purposes of the present application), and the various embodiments formed by such combinations are considered to be part of the present application.

[0204] Example

[0205] The following is a description of exemplary embodiments of the present application in conjunction with the accompanying drawings, including various details of the embodiments of the present application to facilitate understanding. It should be understood that they are considered to be merely exemplary and are by no means intended to limit the scope of protection of the present application. The scope of protection of the present application is limited only by the claims. Therefore, it should be recognized by those of ordinary skill in the art that various changes and modifications can be made to the embodiments described herein without departing from the scope of the present application. Similarly, for clarity and conciseness, the description of well-known functions and structures is omitted in the following description.

[0206] Unless otherwise specified, the reagents and instruments used in the following examples are conventional products that can be obtained commercially. Unless otherwise specified, the experiments were performed according to conventional conditions or conditions recommended by the manufacturer.

[0207] Example 1: Establishment of a screening system for retrotransposon activity

[0208] We designed a high-throughput screening system to detect the insertion activity of different retrotransposons of the present application at specific sites of the 28srRNA gene in the human genome.

[0209] The order of elements in the vector is Figure 1 The specific description is as follows:

[0210] First, Beijing Liuhe Huada Gene Technology Co., Ltd. (BGI TECH SOLUTIONS (BEIJINGLIUHE) CO., LIMITED) was commissioned to synthesize the corresponding DNA sequence and construct a retrotransposon plasmid. The schematic diagram of the example retrotransposon construct is shown in Figure 1 As shown, and from the 5' end to the 3' end there is a 5' untranslated region (5'UTR), a DNA sequence (ORF) encoding an amino acid sequence of a functional protein optimized by human codons, and a 3' untranslated region (3'UTR). The above synthesized sequence is cloned into a plasmid vector pCDNA3.1 that already contains a CMV promoter element and a poly (A) element, and the retrotransposon is transcribed in eukaryotic cells under the control of the CMV promoter, and then translated into a functional protein.

[0211] Secondly, based on the functional characteristics of R2 retrotransposon, we added sequences on both sides of the 5'UTR and 3'UTR of R2 retrotransposon ( Figure 1The added sequences are homologous to the sequences on both sides of the insertion site of the human 28S rRNA gene to improve the integrity of the retrotransposition product (Eickbush DG, Luan DD, Eickbush TH. Integration of Bombyx mori R2 sequences into the 28S ribosomal RNA genes of Drosophila melanogaster. Mol Cell Biol. 2000; 20(1): 213-23.). These two homologous sequences will be transcribed into RNA together during the transcription process of the retrotransposon.

[0212] In order to use a unified method to detect the retrotransposition activity of all retrotransposons, we inserted a consensus sequence (universal seq) between the ORF region encoding the functional protein and the 3'UTR, which includes PCR primer F1 (with the sequence shown in SEQID NO:301, 5'-TGTGCCGAGGCTCAGGCACGCTC-3'), which has high specificity and amplification efficiency in the sequence. After the retrotransposition process is completed and R2 is inserted into the 28s rRNA gene, genomic DNA is extracted and used as a template to amplify the junction of the 28s rRNA gene and the retrotransposon using F1 and reverse primer a1 (with the sequence shown in SEQ ID NO:302, 5'-GGCCTCCCACTTATTCTACACC-3'), about 200bp downstream of the insertion site. Due to the difference in 3'UTR sequence length, PCR products of different lengths will be amplified for different R2 samples. The product is further analyzed by Sanger sequencing to verify whether the directional insertion at the 28s rRNA locus has been achieved.

[0213] Table 1 Functional proteins encoded by retrotransposons and their corresponding untranslated regions

[0214]

[0215]

[0216]

[0217]

[0218] Example 2: Retrotransposition activity assay

[0219] 2.1 Cell treatment (Day 0):

[0220] HEK293T cells (commercially purchased) were cultured to the logarithmic growth phase and then digested with 0.25% trypsin (Thermo) to separate into single cells. 4 Cells were added into a 96-well cell culture plate pre-coated with PDL (Sigma) at 5% CO. 2 , and culture at 37°C overnight.

[0221] 2.2 Cell transfection (Day 1):

[0222] 200 ng of each retrotransposon plasmid constructed in Example 1 was mixed with the transfection reagent Lipofectamine 2000 (Thermo) at a ratio of transfection plasmid mass (μg): transfection reagent volume (μL) = 1:2, and allowed to stand at room temperature for 15 minutes to form a transfection complex. The transfection complex was transferred to a cell culture plate and incubated with cells, and two parallel tests were performed for each sample to be screened.

[0223] 2.3 Cell collection and genome extraction (Day 4)

[0224] 72 hours after transfection, cells were digested and dispersed into single cells with 0.25% trypsin (Thermo). Cells were collected by centrifugation, and genomic DNA of cells was extracted using a genome extraction kit (BEIJING BIOTEKE BIOTECHNOLOGY CO., LTD., DP1202), and the concentration of genomic DNA was measured using a nanodrop one ultra-micro spectrophotometer (Thermo).

[0225] 2.4 Detection

[0226] The genomic DNA of all samples was subjected to PCR analysis according to the conditions in Table 2 and Table 3, and the PCR products were detected by 1% agarose gel electrophoresis.

[0227] Table 2 PCR conditions

[0228]

[0229]

[0230] Table 3 PCR program

[0231]

[0232] 2.5 Test results

[0233] The results of retrotransposition activity are shown in Figure 2 . Figure 2The different lanes in Table 1 correspond to the different retrotransposons numbered 1-100 in Table 1. The results show that the lanes corresponding to most retrotransposons show a single band (e.g., retrotransposons numbered 1-5, 7-12, 14-17, 19-32, 34-92, and 95-100 in Table 1), indicating that these functional proteins encoded by the retrotransposons have certain retrotransposition activity. However, the lanes corresponding to 6 retrotransposons have no bands (e.g., retrotransposons numbered 6, 13, 18, 33, 93, and 94 in Table 1), indicating that these functional proteins encoded by the retrotransposons do not have retrotransposition activity. Compared with these inactive retrotransposons (retrotransposons numbered 6, 13, 18, 33, 93, 94 in Table 1), the other 94 retrotransposons showed better retrotransposition activities (retrotransposons numbered 1-5, 7-12, 14-17, 19-32, 34-92 and 95-100 in Table 1).

[0234] Example 3: Relative quantitative analysis of transposon activity

[0235] Using the human single copy gene 36b4 as an internal reference gene, a relative quantitative analysis of the retrotransposition activity of some retrotransposons in Table 1 was performed. The method is the same as described in Examples 1 and 2, with the only difference being that during the PCR process, an additional PCR primer pair targeting the single copy gene 36b4 was added simultaneously:

[0236] 36b4-F: 5′-CAGCAAGTGGGAAGGTGTAATCC-3′ (sequence shown in SEQ ID NO: 303);

[0237] 36b4-R: 5′-CCCATTCTATCATCAACGGGTACAA-3′ (the sequence is shown in SEQ ID NO: 304).

[0238] The results of retrotransposition activity are shown in Figure 3 The numbers on the different lanes correspond to the corresponding numbered transposon vectors in Table 1. Figure 3 The detection results in the quantitative analysis were performed using Image Lab 6.1.0 (Bio-Rad Laboratories Inc.) software. The calculation formula = gray value of R2 band / gray value of 36b4 band. The quantitative analysis results are shown in Figure 4 and Table 4. The above results show that all functional proteins encoded by the retrotransposon in the present application have certain retrotransposition activity, and some functional proteins have relatively high activity.

[0239] Table 4 Relative quantitative results of retrotransposon activity

[0240]

[0241]

[0242] Example 4: Detection of the 5' junction of retrotransposons

[0243] Method for detecting the 5' junction of the 28s rRNA gene and the retrotransposon in Table 1, which shows the integrity of the retrotransposition event. The method is the same as described in Examples 1 and 2, with the only difference being that three reverse primers are designed for different positions in the transposon ORF region and 5'UTR sequence (the sequences of the reverse primers are shown in Table 4), and are paired with a forward primer (28s-up-F1, sequence as shown in SEQ ID NO:305) about 200 bp upstream of the 28s rRNA gene insertion site for PCR analysis. The integrity of the 5' end sequences of these samples after insertion is tested to preliminarily determine the retrotransposition integrity of different R2 samples.

[0244] The results of the retrotransposition integrity test were as follows Figure 5 As shown. The numbers on different lanes correspond to the transposon vectors with corresponding numbers in Table 1. The three lanes of each vector correspond to the PCR bands obtained by pairing three reverse primers (having the sequences shown in Table 5) with a forward primer (28s-up-F1, the sequence is shown in SEQ ID NO:305). The results show that the retrotransposition product obtained from the functional protein encoded by the retrotransposon in the present application has relatively good integrity.

[0245] Table 5 Primer sequences used for retrotransposition integrity determination

[0246]

[0247]

[0248]

[0249]

[0250] It should be noted that the above are only preferred examples of the present application and are not intended to limit the present application. For those of ordinary skill in the art, various modifications and changes can be made to the present application. Although specific embodiments have been described, for the applicant or other persons skilled in the art, there may be or are currently unable to foresee substitutions, modifications, changes, improvements and substantial equivalents of the above-described embodiments. Therefore, the attached claims submitted and the claims that may be modified are intended to cover all such substitutions, modifications, changes, improvements and substantial equivalents. It is important that with the evolution of technology, many elements described herein can be replaced by equivalent elements that appear after the present application.

Claims

1. An isolated functional protein encoded by a retrotransposon, wherein the functional protein has a functional protein sequence selected from the following (i) or a variant sequence of the aforementioned functional protein having functional protein activity among (ii) to (iv): (i) at least one of the amino acid sequences shown in any one of SEQ ID NOs: 1-94; (ii) at least one of the sequences obtained by deleting, substituting, inserting or mutating 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids in the amino acid sequence shown in any one of SEQ ID NOs: 1-94; (iii) at least one of the amino acid sequences having at least 70%, 80%, 90%, 95% or 99% identity to the amino acid sequence shown in any one of SEQ ID NOs: 1-94; and (iv) at least one of the sequences obtained by further fusion of the amino acid sequence represented by any one of SEQ ID NOs: 1-94 with other sequences.

2. An isolated functional protein encoded by a retrotransposon, wherein the functional protein comprises an amino acid sequence represented by formula (I): C(X1) a C(X2) b H(X3) c H(I), in, a, b, and c are the number of amino acids; C is cysteine; H is histidine; (X1) is any amino acid, and a is 1, 2, 3 or 4; (X2) is any amino acid, and b is 11, 12, 13, 14, 15, 16 or 17; and (X3) is any amino acid, and c is 4.

3. An isolated functional protein encoded by a retrotransposon, wherein the functional protein comprises an amino acid sequence represented by formula (II): G(X4) d Q G D(X5) e S(X6) f F(X7) g D(II), in, d, e, f, and g are the number of amino acids; D is aspartic acid; F is phenylalanine; G is glycine; Q is glutamine; S is serine; (X4) is any amino acid, and d is 2; (X5) is any amino acid, and e is 2; (X6) is any amino acid, and f is 3; and (X7) is any amino acid, and g is 30, 31, 32, 33, 34, 35 or 36.

4. An isolated functional protein encoded by a retrotransposon, wherein the functional protein comprises an amino acid sequence represented by formula (III): C(X8) h C(X9) i E(X 10 ) j H(X 11 ) k C(X 12 ) l R H(X 13 ) m P D(X 14 ) n (X 15 )(X 16 ) o K(X 17 ) p Y(III), in, h, i, j, k, l, m, n, o, and p are the numbers of amino acids; C is cysteine; D is aspartic acid; E is glutamate; H is histidine; K is lysine; R is arginine; P is proline; Y is tyrosine; (X8) is any amino acid, and h is 2, 3 or 4; (X9) is any amino acid, and i is 3, 4, 5, 6, 7, 8, 9 or 10; (X 10 ) is any amino acid, and j is 3; (X 11 ) is any amino acid, and k is 4; (X 12 ) is any amino acid, and l is 9; (X 13 ) is any amino acid, and m is 31, 32, 33, 34, 35 or 36; (X 14 ) is any amino acid, and n is 11, 12, 13, 14, 15, 16, 17, 18 or 19; (X 15 ) is aspartic acid or glutamic acid; (X 16 ) is any amino acid, and o is 15, 16, 17, 18 or 19; and (X 17 ) is any amino acid, and p is 3.

5. An isolated functional protein encoded by a retrotransposon, wherein the functional protein comprises at least two of the amino acid sequences represented by formula (I), formula (II) and formula (III).

6. An isolated functional protein encoded by a retrotransposon, wherein the functional protein comprises the amino acid sequences shown in formula (I), formula (II) and formula (III).

7. The functional protein according to any one of claims 1 to 6, wherein the retrotransposon is a non-LTR (non-long terminal repeat) retrotransposon.

8. The functional protein according to claim 7, wherein the non-LTR retrotransposon is a restriction enzyme-like nuclease (RLE) retrotransposon.

9. The functional protein according to any one of claims 1 to 8, wherein the retrotransposon is an R2 family retrotransposon.

10. The functional protein according to any one of claims 1-9, wherein the functional protein comprises a DNA binding domain capable of binding to a specific region in a cell genome, preferably, the DNA binding domain is capable of binding to a gene encoding 28s rRNA.

11. The functional protein according to claim 10, wherein the functional protein further comprises an RNA binding domain, a reverse transcriptase functional domain and / or a nuclease functional domain. 12 . The functional protein according to claim 1 , wherein the species origin of the functional protein includes Arthropoda or Chordata. 13 . The functional protein according to claim 12 , wherein the species of the functional protein include Insecta, Actinopterygii, Chondrichthyes, Testudinata, Lepidosauria or Aves.

14. The functional protein according to claim 13, wherein the species of the functional protein include goshawk (Accipiter gentilis), grouper turtle (Actinemys marmorata), tri-colored blackbird (Agelaius tricolor), noctuidae (Agrochola macilenta), rice louse wasp (Anagrus nilaparvatae), red-tailed mining bee (Andrena haemorrhoa), spider hunting wasp (Anoplius nigerrimus), Bai's desert bunting (Artemisiospiza belli), open-armed anti-jawed braconid wasp (Asobara japonica), yellow-winged sawfly (Athaliarosae), mole moth (Blastobasis lacticolella), long-cheeked bumblebee (Bombus hortorum), sleeping bumblebee (Bombus hypnorum), early bumblebee (Bombus pratorum), Vancouver bumblebee (Bombusvancouverensis), Ino small leopard butterfly (Brenthis ino), Cardiocondylaobscurior, Cerceris rybyensis, Clusia tigrina, Colaptes auratus, Crematogaster levior, Crotalus tigris, Cuora mccordi, Dinocampus coccinellae, Dolichovespula saxonica, Drosophila albomicans, Drosophilasaltans, Ennomos quercinarius, Eopsaltria australis, Eristalis pertinax, Erithacus rubecula, Euphyes dion, Heliconius hecale), Hesperophylax magnus, Ichneumonxanthorius, Jera tricuspidata, Junonia litoralis, Lasioglossum baleicum, Lasioglossumlativentre), Lasioglossummorio, Leptopilina heterotoma, Lysandra coridon, Marasmarcha lunaedactyla, Mimumesa dahlbomi, Muschampia plurimacula, Neodiprion fabricii, Neodiprion pinetum, Nephrotoma flavescens, Nomada fabriciana, Nylanderia fulva, Nymphalis io, Papilio machaon, Pararge aegeria, Podocnemis expansa, Poecilia wingei, Prinia pygmy subflava), Scaptomyza hsui, Schistocerca americana, Seladonia tumulorum, Sphecodes monilicornis, Tibetan loach, Trypoxylus dichotomus, Urbanus tucuti, Venturia canescens, Vespa crabro, Zaprionus camerounensis, or Zaprionus kolodkinae.

15. A nucleic acid, wherein the nucleic acid encodes the functional protein according to any one of claims 1-14.

16. A nucleic acid set comprising a 5' untranslated region, wherein the 5' untranslated region comprises at least one of the nucleotide sequences shown in SEQ ID NOs: 95-188.

17. A nucleic acid set comprising a 3' untranslated region, wherein the 3' untranslated region comprises at least one of the nucleotide sequences shown in SEQ ID NOs: 189-282.

18. A nucleic acid group comprising a 5' untranslated region and a 3' untranslated region, wherein the 5' untranslated region comprises a nucleotide sequence shown in any one of SEQ ID NOs: 95-188 or a variant thereof, and the 3' untranslated region comprises a nucleotide sequence shown in any one of SEQ ID NOs: 189-282 or a variant thereof, and the RNA transcribed from the nucleic acid group is capable of binding to a functional protein encoded by a specific retrotransposon.

19. A nucleic acid construct comprising the nucleic acid according to claim 15 and / or the nucleic acid group according to any one of claims 16 to 18.

20. nucleic acid construct according to claim 19, wherein said nucleic acid construct also comprises exogenous nucleic acid fragment, and said exogenous nucleic acid fragment is operably inserted into the nucleic acid construct through the multiple cloning insertion site, and said exogenous nucleic acid fragment can have one or more, can be identical or different.

21. The nucleic acid construct according to claim 20, wherein the exogenous nucleic acid fragment comprises any gene of interest or any gene capable of being transposed, preferably, the exogenous nucleic acid fragment comprises a natural functional protein gene, an artificial chimeric gene, and / or a non-coding RNA gene.

22. The nucleic acid construct of claim 21, wherein the native functional protein gene comprises a fluorescence-based reporter gene, a luciferase gene, and / or an antibiotic resistance gene.

23. The nucleic acid construct of claim 21, wherein the artificial chimeric gene comprises a chimeric antigen receptor gene.

24. The nucleic acid construct of claim 21, wherein the non-coding RNA gene comprises rRNA, tRNA, small interfering RNA (siRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), microRNA (miRNA), and / or long non-coding RNA (lncRNA).

25. The nucleic acid construct of claim 19, wherein the nucleic acid construct further comprises a promoter and a poly (A) sequence.

26. The nucleic acid construct of claim 25, wherein the promoter comprises CMV, EF1a, SV40, PGK, UbC, human beta actin, CAG, TRE, UAS, Ac5, GFAP, polyhedrin promoter, TBG, ALB, ApoEHCR-hAAT, CaMKIIa, GAL1, TEF1, GDS, ADH1, CaMV35S, Ubi, H1, U6, T7, T7lac, Sp6, araBAD, trp, lac, Ptac, or pL.

27. A composition, wherein the composition comprises: A functional protein or a functional fragment thereof encoded by an R2 family retrotransposon, or a nucleic acid encoding the functional protein or a functional fragment thereof, wherein the functional protein or the functional fragment thereof has the function of catalyzing the insertion of an exogenous nucleic acid fragment into a cell genome; and A nucleic acid group, wherein RNA transcribed from the nucleic acid group can be recognized by a functional protein encoded by a specific retrotransposon or a functional fragment thereof.

28. The composition according to claim 27, wherein the composition is selected from at least one of the following groups (1) to (95), any one of the following groups (1) to (94) comprising: a functional protein-related sequence and a nucleic acid group, (1) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 1 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 95; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 189; (2) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 2 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 96; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 190; (3) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 3 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 97; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 191; (4) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 4 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 98; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 192; (5) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 5 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 99; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 193; (6) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 6 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 100; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 194; (7) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 7 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 101; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 195; (8) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 8 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 102; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 196; (9) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 9 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 103; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 197; (10) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 10 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 104; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 198; (11) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 11 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 105; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 199; (12) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 12 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 106; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 200; (13) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 13 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 107; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 201; (14) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 14 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 108; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 202; (15) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 15 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 109; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 203; (16) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 16 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 110; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 204; (17) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 17 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 111; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 205; (18) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 18 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 112; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 206; (19) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 19 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 113; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 207; (20) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 20 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 114; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 208; (21) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 21 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 115; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 209; (22) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 22 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 116; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 210; (23) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 23 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 117; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 211; (24) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 24 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 118; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 212; (25) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 25 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 119; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 213; (26) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 26 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 120; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 214; (27) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 27 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 121; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 215; (28) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 28 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 122; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 216; (29) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 29 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 123; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 217; (30) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 30 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 124; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 218; (31) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 31 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 125; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 219; (32) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 32 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 126; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 220; (33) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 33 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 127; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 221; (34) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 34 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 128; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 222; (35) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 35 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 129; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 223; (36) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 36 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 130; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 224; (37) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 37 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 131; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 225; (38) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 38 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 132; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 226; (39) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 39 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 133; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 227; (40) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 40 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 134; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 228; (41) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 41 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 135; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 229; (42) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 42 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 136; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 230; (43) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 43 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 137; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 231; (44) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 44 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 138; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 232; (45) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 45 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 139; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 233; (46) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 46 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 140; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 234; (47) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 47 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 141; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 235; (48) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 48 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 142; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 236; (49) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 49 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 143; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 237; (50) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 50 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 144; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 238; (51) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 51 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 145; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 239; (52) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 52 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 146; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 240; (53) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 53 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 147; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 241; (54) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 54 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 148; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 242; (55) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 55 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 149; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 243; (56) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 56 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 150; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 244; (57) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 57 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 151; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 245; (58) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 58 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 152; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 246; (59) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 59 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 153; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 247; (60) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 60 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 154; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 248; (61) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 61 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 155; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 249; (62) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 62 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 156; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 250; (63) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 63 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 157; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 251; (64) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 64 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 158; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 252; (65) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 65 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 159; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 253; (66) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 66 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 160; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 254; (67) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 67 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 161; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 255; (68) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 68 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 162; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 256; (69) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 69 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 163; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 257; (70) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 70 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 164; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 258; (71) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 71 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 165; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 259; (72) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 72 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 166; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 260; (73) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 73 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 167; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 261; (74) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 74 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 168; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 262; (75) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 75 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 169; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 263; (76) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 76 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 170; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 264; (77) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 77 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 171; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 265; (78) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 78 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 172; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 266; (79) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 79 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 173; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 267; (80) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 80 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 174; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 268; (81) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 81 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 175; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 269; (82) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 82 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 176; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 270; (83) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 83 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 177; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 271; (84) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 84 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 178; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 272; (85) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 85 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 179; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 273; (86) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 86 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 180; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 274; (87) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 87 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 181; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 275; (88) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 88 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 182; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 276; (89) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 89 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 183; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 277; (90) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 90 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 184; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 278; (91) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 91 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 185; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 279; (92) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 92 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 186; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 280; (93) The functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 93 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 187; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 281; (94) the functional protein-related sequence is an amino acid sequence comprising the sequence shown in SEQ ID NO: 94 or a nucleic acid encoding the amino acid sequence; the nucleic acid group comprises a 5' untranslated region and a 3' untranslated region, the 5' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 188; the 3' untranslated region is a nucleotide sequence comprising the sequence shown in SEQ ID NO: 282; or (95) A variant of any one of the aforementioned groups (1) to (94), in, The functional protein-related sequence is an amino acid sequence of a variant of each group of functional proteins or a nucleic acid sequence encoding the variant, wherein the variant has a variant sequence of the aforementioned functional protein having functional protein activity selected from the following (i)-(iii): (i) at least one of the sequences obtained by deleting, replacing, inserting or mutating 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids in the amino acid sequence of each group of functional proteins; (ii) at least one of the amino acid sequences that is at least 70%, 80%, 90%, 95% or 99% identical to the amino acid sequence shown in any one of SEQ ID NOs: 1-94; as well as (iii) at least one of the sequences obtained by further fusion of the amino acid sequence represented by any one of SEQ ID NOs: 1-94 with other sequences.

29. The composition according to any one of claims 27 or 28, wherein the nucleic acid and / or nucleic acid group further comprises a promoter and a poly (A) sequence.

30. The composition of claim 29, wherein the promoter comprises CMV, EF1a, SV40, PGK, UbC, human beta actin, CAG, TRE, UAS, Ac5, GFAP, polyhedrin promoter, TBG, ALB, ApoEHCR-hAAT, CaMKIIa, GAL1, TEF1, GDS, ADH1, CaMV35S, Ubi, H1, U6, T7, T7lac, Sp6, araBAD, trp, lac, Ptac, or pL.

31. The composition of any one of claims 27 or 28, wherein the set of nucleic acids further comprises exogenous nucleic acid fragments.

32. The composition according to claim 31, wherein the exogenous nucleic acid fragment is operably inserted into the nucleic acid group through a multiple cloning insertion site, and the exogenous nucleic acid fragment may be one or more and may be the same or different.

33. The composition of claim 32, wherein the exogenous nucleic acid fragment comprises any gene of interest or any gene capable of being transposed, preferably, the exogenous nucleic acid fragment comprises a natural functional protein gene, an artificial chimeric gene, and / or a non-coding RNA gene.

34. The composition of claim 33, wherein the native functional protein gene comprises a fluorescence-based reporter gene, a luciferase gene, and / or a resistance gene.

35. The composition of claim 33, wherein the artificial chimeric gene comprises a chimeric antigen receptor gene.

36. The composition of claim 33, wherein the non-coding RNA gene comprises rRNA, tRNA, small interfering RNA (siRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), micro RNA (miRNA), and / or long non-coding RNA (lncRNA).

37. A recombinant vector, wherein the recombinant vector comprises a nucleic acid encoding a functional protein according to any one of claims 1-14, a nucleic acid according to claim 15, a nucleic acid group according to any one of claims 16-18, a nucleic acid construct according to any one of claims 19-26, or a composition according to any one of claims 27-36.

38. The recombinant vector of claim 37, wherein the recombinant vector comprises a recombinant cloning vector, a recombinant eukaryotic expression plasmid, or a recombinant viral vector.

39. The recombinant vector of claim 38, wherein the recombinant eukaryotic expression plasmid comprises pcDNA3.1, pCMV, pUC18, pUC19, pUC57, pBAD, pET, pENTR, pGenlenti, or pAAV.

40. The recombinant vector of claim 38, wherein the recombinant viral vector comprises a recombinant adenoviral vector, a recombinant adeno-associated viral vector, a recombinant retroviral vector, a recombinant herpes simplex viral vector, or a recombinant vaccinia viral vector.

41. A recombinant host cell, wherein the recombinant host cell comprises a functional protein according to any one of claims 1-14, a nucleic acid according to claim 15, a nucleic acid group according to any one of claims 16-18, a nucleic acid construct according to any one of claims 19-26, a composition according to any one of claims 27-36, or a recombinant vector according to any one of claims 37-40.

42. The recombinant host cell of claim 41, wherein the recombinant host cell comprises an animal cell, a plant cell, an algae cell, a fungal cell, a yeast cell, or a bacterial cell.

43. The recombinant host cell of claim 42, wherein the animal cell comprises a mammalian cell.

44. The recombinant host cell of claim 43, wherein the mammalian cell comprises primary cells (e.g., mesenchymal stem cells, endothelial cells, epithelial cells, fibroblasts, keratinocytes, melanocytes, smooth muscle cells, and immune cells), immortalized cell lines (e.g., HEK293, NIH-3T3, RAW-264.7, STO, VERO, CT26, hTERT immortalized human endothelial / epithelial / fibroblast / keratinocyte / ductal / cell lines), cancer cell lines (e.g., Hela, HepG2 / 3, HL-60, HT-1080, HT-29 , A549, SW620, HCT-15, HCT116, MDA-MB-231, MCF7, SK-OV-3, PANC-1, AsPc-1, THP-1, Huh7, KG-1, RAJI, HB-CB, Jurkat, K562, CRL5826, CHO, MDCK, Renca), embryonic stem cell lines (e.g., H1, H9, WIBR2, WIBR3, G-Olig2, ESF158, RW.4, R1, D3) and differentiated cells thereof, or induced pluripotent stem cell lines and differentiated cells thereof.

45. A method for introducing an exogenous nucleic acid fragment into a host cell genome, wherein the method comprises: The functional protein according to any one of claims 1 to 14, the nucleic acid according to claim 15, the nucleic acid group according to any one of claims 16 to 18, the nucleic acid construct according to any one of claims 19 to 26, the composition according to any one of claims 27 to 36, or the recombinant vector according to any one of claims 37 to 40 is delivered into the host cell.

46. ​​A method for editing a host cell genome, wherein the method comprises: The functional protein according to any one of claims 1 to 14, the nucleic acid according to claim 15, the nucleic acid group according to any one of claims 16 to 18, the nucleic acid construct according to any one of claims 19 to 26, the composition according to any one of claims 27 to 36, or the recombinant vector according to any one of claims 37 to 40 is delivered into the host cell.

47. A method for obtaining a host cell comprising an exogenous nucleic acid fragment in its genome, wherein the method comprises: The functional protein according to any one of claims 1 to 14, the nucleic acid according to claim 15, the nucleic acid group according to any one of claims 16 to 18, the nucleic acid construct according to any one of claims 19 to 26, the composition according to any one of claims 27 to 36, or the recombinant vector according to any one of claims 37 to 40 is delivered into the host cell.

48. The method of any one of claims 45-47, wherein the delivery method comprises cationic liposome delivery, lipid nanoparticle delivery, cationic polymer delivery, vesicle-exosome delivery, gold nanoparticle delivery, polypeptide and protein delivery, retroviral delivery, lentiviral delivery, adenoviral delivery, adeno-associated virus delivery, electroporation delivery, Agrobacterium infection delivery, or gene gun delivery.

49. The method of any one of claims 45-47, wherein the host cell comprises an animal cell, a plant cell, an algae cell, a fungal cell, a yeast cell, or a bacterial cell.

50. The method of claim 49, wherein the host cell comprises a mammalian cell.

51. The method of claim 50, wherein the host cell comprises a primary cell (e.g., mesenchymal stem cells, endothelial cells, epithelial cells, fibroblasts, keratinocytes, melanocytes, smooth muscle cells, immune cells), an immortalized cell line (e.g., HEK293, NIH-3T3, RAW-264.7, STO, VERO, CT26, hTERT immortalized human endothelial / epithelial / fibroblast / keratinocyte / ductal / cell line), a cancer cell line (e.g., Hela, HepG2 / 3, HL-60, HT-1080, HT-29, A5 49, SW620, HCT-15, HCT156, MDA-MB-231, MCF7, SK-OV-3, PANC-1, AsPc-1, THP-1, Huh7, KG-1, RAJI, HB-CB, Jurkat, K562, CRL5826, CHO, MDCK, Renca), embryonic stem cell lines (e.g., H1, H9, WIBR2, WIBR3, G-Olig2, ESF158, RW.4, R1, D3) and differentiated cells thereof, or induced pluripotent stem cell lines and differentiated cells thereof.

52. Use of the functional protein according to any one of claims 1-14, the nucleic acid according to any one of claims 15, the nucleic acid group according to any one of claims 16-18, the nucleic acid construct according to any one of claims 19-26, the composition according to any one of claims 27-36, the recombinant vector according to any one of claims 37-40, or the recombinant host cell according to any one of claims 41-44 in introducing exogenous nucleic acid fragments into the host cell genome.

53. The use of claim 52, wherein the host cell comprises an animal cell, a plant cell, an algae cell, a fungal cell, a yeast cell, or a bacterial cell.

54. The use of claim 53, wherein the host cell comprises a mammalian cell.

55. The use according to claim 54, wherein the host cell comprises primary cells (e.g., mesenchymal stem cells, endothelial cells, epithelial cells, fibroblasts, keratinocytes, melanocytes, smooth muscle cells, immune cells), immortalized cell lines (e.g., HEK293, NIH-3T3, RAW-264.7, STO, VERO, CT26, hTERT immortalized human endothelial / epithelial / fibroblast / keratinocyte / ductal / cell lines), cancer cell lines (e.g., Hela, HepG2 / 3, HL-60, HT-1080, HT-29, A5 49, SW620, HCT-15, HCT156, MDA-MB-231, MCF7, SK-OV-3, PANC-1, AsPc-1, THP-1, Huh7, KG-1, RAJI, HB-CB, Jurkat, K562, CRL5826, CHO, MDCK, Renca), embryonic stem cell lines (e.g., H1, H9, WIBR2, WIBR3, G-Olig2, ESF158, RW.4, R1, D3) and differentiated cells thereof, or induced pluripotent stem cell lines and differentiated cells thereof.

56. Use of the functional protein according to any one of claims 1-14, the nucleic acid according to claim 15, the nucleic acid group according to any one of claims 16-18, the nucleic acid construct according to any one of claims 19-26, the composition according to any one of claims 27-36, the recombinant vector according to any one of claims 37-40, or the recombinant host cell according to any one of claims 41-44 in the preparation of drugs or preparations for gene therapy, cell therapy, genome research, or stem cell induction and post-induced differentiation.

57. A kit, wherein the kit comprises a functional protein according to any one of claims 1-14, a nucleic acid according to claim 15, a nucleic acid set according to any one of claims 16-18, a nucleic acid construct according to any one of claims 19-26, a composition according to any one of claims 27-36, a recombinant vector according to any one of claims 37-40, or a recombinant host cell according to any one of claims 41-44.

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