Drug delivery composition

Abstract

A drug delivery composition comprising acid-resistant cells encapsulating a drug therein. Additionally, an acid-resistant cell which is an acid-resistant cell encapsulating a drug therein, and the drug is locally present in a pouch-like membrane structure possessed by the acid-resistant cell.

Description

Technical Field

[0001] The present invention relates to a drug delivery composition; in addition, it also relates to acid-resistant cells, a drug carrier and a method for preparing the acid-resistant cells that can be used for the drug delivery composition.

[0002] This application claims priority based on Japanese Patent Application No. 2019-069029 filed in Japan on March 29, 2019, the contents of which are incorporated herein by reference. Background Art

[0003] In animals with digestive tracts, such as humans, food ingested through the mouth is delivered to the stomach via the esophagus. For example, when administering drugs orally, particularly when the drugs contain peptides or proteins as their primary ingredients, there is a high likelihood that the drugs will be broken down by enzymes in the stomach. Furthermore, because the stomach is highly acidic, even low-molecular-weight drugs may be non-enzymatically broken down in the stomach. Furthermore, even when acidic compounds are intended to be absorbed by the intestines, they are sometimes absorbed by the stomach. Therefore, oral administration using capsules that do not dissolve in the stomach but dissolve in the intestines is useful.

[0004] As methods for achieving intestinal drug delivery, known technologies include a technology called bilosome that utilizes the property of a substance with protein B introduced into lipids to be stable in the stomach (non-patent document 1), a rice vaccine that utilizes the resistance of protein bodies, which are organelles of rice, to digestive enzymes (non-patent document 2), and a spore vaccine that utilizes spores that are resistant to digestive enzymes, temperature changes, and pH changes (non-patent document 3).

[0005] As vaccines aiming for industrial oral vaccines, vaccines utilizing yeast are known. For example, Patent Document 1 describes an oral vaccine in which antigenic proteins are expressed within yeast cells. Patent Document 1 shows that freeze-drying the yeast cells prevents digestion in the stomach and jejunum but allows for digestion and decomposition in the ileum. However, the release of antigenic proteins from yeast depends on the function of digestive enzymes in the small intestine. Patent Document 2 suggests the use of transmucosal / oral administration of yeast strains harboring foreign genes to induce immunity, but also states that the proteins from the yeast used are also antigenic.

[0006] Prior art literature

[0007] Patent Literature

[0008] Patent Document 1: International Publication No. 2006 / 028214

[0009] Patent Document 2: Japanese Patent Application No. 2012-508697

[0010] Non-patent literature

[0011] Non-patent document 1: Mann JF et al., Lipid vesicle size of an oral influenza vaccine delivery vehicle influences the Th1 / Th2 bias in the immune response and protection against infection. Vaccine. 2009 Jun 2; 27(27): 3643-9.

[0012] Non-patent document 2: Nochi T et al., Rice-based mucosal vaccination as a global strategy for cold-chain-and needle-free vaccination. Proc Natl Acad Sci USA. 2007 Jun 26; 104(26): 10986-91.

[0013] Non-patent document 3: Huang JM et al., Mucosal delivery of antigens using adsorption to bacterial spores. Vaccine. 2010 Jan 22; 28(4): 1021-30. Summary of the Invention

[0014] Problems to be solved by the invention

[0015] In animal husbandry, if an infectious disease occurs, it is difficult to suppress the spread of infection, and sometimes a large number of livestock are killed. Among infectious diseases, there are also infectious diseases that are believed to be preventable through intestinal immunity. It is an urgent issue to develop a technology for establishing immunity to pathogens in the intestines of livestock. In addition, by imparting intestinal immunity, there is the possibility of imparting other mucosal immunity and systemic immunity. Therefore, it is required to develop an enteric-coated composition that can be directly delivered to the intestine by orally administering a vaccine. However, the technologies described in Non-Patent Documents 1 to 3 have cost issues when used in animal husbandry.

[0016] Therefore, an object of the present invention is to provide a novel drug delivery composition capable of delivering drugs to the intestine, acid-resistant cells and drug carriers that can be used in the drug delivery composition, and a method for producing the acid-resistant cells.

[0017] Means used to solve problems

[0018] The present invention includes the following technical solutions.

[0019] (1) A drug delivery composition comprising acid-resistant cells encapsulating a drug.

[0020] (2) The drug delivery composition according to (1), wherein the drug is locally present in the bag-like membrane structure possessed by the acid-resistant cells.

[0021] (3) The drug delivery composition according to (2), wherein the bag-like membrane structure is at least one selected from the group consisting of exogenous liposomes and cell organelles.

[0022] (4) The drug delivery composition according to (3), wherein the cell organelle is at least one selected from the group consisting of mitochondria, chloroplasts, endoplasmic reticulum, vacuoles, nuclei, peroxisomes, and Golgi apparatus.

[0023] (5) The drug delivery composition according to any one of (1) to (4), wherein the drug is at least one selected from the group consisting of low molecular weight compounds, peptides, proteins, and nucleic acids.

[0024] (6) The drug delivery composition according to any one of (1) to (5), wherein the drug is a drug that acts in the intestine.

[0025] (7) The drug delivery composition according to any one of (1) to (6), wherein the drug is an immunogenic drug.

[0026] (8) The drug delivery composition according to any one of (1) to (7), wherein the acid-resistant cells are cells that undergo cell rupture under conditions of pH 7 or higher.

[0027] (9) The drug delivery composition according to any one of (1) to (8), wherein the acid-resistant cells are cells resistant to acidic conditions of pH 1 to 3.

[0028] (10) The drug delivery composition according to any one of (1) to (9), wherein the acid-resistant cells are cells of algae belonging to the class Cyanidiophyceae.

[0029] (11) A feed comprising the drug delivery composition according to any one of (1) to (10).

[0030] (12) A pharmaceutical product comprising the drug delivery composition according to any one of (1) to (10).

[0031] (13) An acid-resistant cell containing a drug.

[0032] (14) The acid-resistant cell according to (13), wherein the drug is locally present in a bag-like membrane structure possessed by the acid-resistant cell.

[0033] (15) The acid-resistant cell according to (13), wherein the drug is locally present inside or outside the bag-like membrane structure of the acid-resistant cell.

[0034] (16) The acid-resistant cell according to any one of (13) to (15), wherein the drug is at least one selected from the group consisting of low molecular weight compounds, peptides, proteins, and nucleic acids.

[0035] (17) A method for producing acid-resistant cells as described in (14), which comprises the step of introducing a gene encoding a fusion protein into the acid-resistant cells, wherein the fusion protein contains a peptide or protein as a drug and a peptide or protein that is locally present relative to a cell membrane or organelle.

[0036] In addition, the present invention also includes the following technical solutions.

[0037] (18) A drug carrier comprising acid-resistant cells.

[0038] (19) The drug carrier according to (18), wherein the acid-resistant cells are cells that undergo cell rupture at a pH of 7 or above.

[0039] (20) The drug carrier according to (18) or (19), wherein the acid-resistant cells are cells resistant to acidic conditions of pH 1 to 3.

[0040] (21) The drug carrier according to any one of (18) to (20), wherein the acid-resistant cells are cells of algae belonging to the Cyanidiophyceae.

[0041] (22) A drug capsule comprising a drug encapsulated in the drug carrier according to any one of (18) to (21).

[0042] (23) The drug carrier according to (22), wherein the drug is locally present in the bag-like membrane structure possessed by the acid-resistant cells.

[0043] In addition, the present invention also includes the following technical solutions.

[0044] (24) An acid-resistant cell comprising an exogenous substance.

[0045] (25) The acid-resistant cell according to (24), wherein the exogenous substance is locally present in a bag-like membrane structure possessed by the acid-resistant cell.

[0046] (26) The acid-resistant cell according to (24) or (25), wherein the exogenous substance is at least one selected from the group consisting of low molecular weight compounds, peptides, proteins, nucleic acids, and synthetic high molecular weight compounds.

[0047] (27) The acid-resistant cell according to any one of (24) to (26), wherein the exogenous substance is a substance that acts in the intestine.

[0048] (28) The acid-resistant cell according to any one of (24) to (27), wherein the exogenous substance is an immunogenic substance.

[0049] (29) The acid-resistant cell according to any one of (25) to (28), wherein the bag-like membrane structure is at least one selected from the group consisting of exogenous liposomes, cell membranes, and organelles.

[0050] (30) The acid-resistant cell according to (29), wherein the organelle is at least one selected from the group consisting of mitochondria, chloroplasts, endoplasmic reticulum, vacuoles, nuclei, peroxisomes, and Golgi apparatus.

[0051] (31) The acid-resistant cell according to any one of (24) to (30), which is a cell that undergoes cell rupture under conditions of pH 7 or higher.

[0052] (32) The acid-resistant cell according to any one of (24) to (31), which is a cell resistant to acidic conditions of pH 1 to 3.

[0053] (33) The acid-resistant cell according to any one of (24) to (32), which is a cell of algae belonging to the class Cyanidiophyceae.

[0054] (34) A feed comprising the acid-resistant cells according to any one of (24) to (33).

[0055] (35) A pharmaceutical product comprising the acid-resistant cells according to any one of (24) to (33).

[0056] (36) A method for administering the exogenous substance, comprising orally administering the acid-resistant cells according to any one of (24) to (33) to a subject.

[0057] (37) A method for raising an animal, comprising feeding the animal with the acid-resistant cells according to any one of (24) to (33).

[0058] (38) A method for imparting intestinal immunity, comprising orally administering the acid-resistant cells according to any one of (24) to (33).

[0059] Effects of the Invention

[0060] The present invention can provide a novel drug delivery composition capable of delivering drugs to the intestine, acid-resistant cells and drug carriers that can be used in the drug delivery composition, and a method for producing the acid-resistant cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0061] Figure 1 The figure shows the results of immunoblotting using an anti-GFP antibody for GAPDH-GP-sfGFP expressing strains cultured in the presence and absence of MG-132. In the figure, the arrow indicates the band of the GAPDH-GP-sfGFP protein.

[0062] Figure 2 Fluorescence microscopy images of a GAPDH-GP-sfGFP expression strain: (A) PC: Phase contrast microscopy image showing cell outlines; (B) Chl: Autofluorescence image of chloroplasts; (C) sfGFP: Fluorescence image of sfGFP.

[0063] Figure 3 This is a diagram showing the structure of the DNA fragment used for preparing the Chl-TP-3HA-GP-Co1 expression strain in Example 2.

[0064] Figure 4 The graph shows the results of immunoblotting using anti-HA antibodies for Chl-TP-3HA-GP-Co1 expression strains cultured in the presence and absence of MG-132. In the graph, the arrow indicates the band of the Chl-TP-3HA-GP-Co1 protein.

[0065] Figure 5 Fluorescence microscopy images of the Chl-TP-3HA-GP-Co1 expression strain. (A) PC: Phase contrast microscopy image showing cell outlines; (B) Chl: Autofluorescence image of chloroplasts; (C) sfGFP: Immunofluorescence staining image using an anti-HA antibody.

[0066] Figure 6 Figures show the results of immunoblotting to evaluate anti-GP protein antibody production in mice administered a suspension of the sfGFP-expressing strain (control suspension-administered group), a suspension of the Chl-TP-3HA-GP-Col-expressing strain (suspension-administered group), or an alginate-immobilized diet of the Chl-TP-3HA-GP-Col-expressing strain (alginate-immobilized diet-administered group). (A): Alginate-immobilized diet-administered group; (B): Suspension-administered group; (C): Control suspension-administered group. Numbers 1 to 4 indicate individual mouse numbers.

[0067] Figure 7 A molecular phylogenetic tree of algae belonging to the Hot Spring Rhodophyceae, based on the chloroplast ribulose 1,5-bisphosphate carboxylase / oxygenase large subunit gene, is shown. Local bootstrap values ​​based on maximum likelihood (only those above 50 are shown on the left) and posterior probabilities based on Bayesian methods (only those above 0.95 are shown on the right) are shown near each branch. The known species Cyanidioschyzon merolae (a hot spring red alga) is enclosed by a dotted line, while strains YFU3 and HKN1 are enclosed by a solid line. DETAILED DESCRIPTION

[0068] [definition]

[0069] In this specification, the terms "peptide" and "protein" are used interchangeably to refer to polymers of amino acids bound by amide bonds. A "peptide" or "protein" may be a polymer of natural amino acids, a polymer of natural amino acids and non-natural amino acids (chemical analogs, modified derivatives, etc. of natural amino acids), or a polymer of non-natural amino acids. Unless otherwise specified, amino acid sequences are listed from the N-terminal end to the C-terminal end.

[0070] There is no particular limitation on the number of amino acid residues constituting a "peptide" or "protein," and amino acid polymers having two or more amino acid residues are also included in "peptides" or "proteins." In this specification, unless otherwise specified, polymers having a large number of amino acid residues (e.g., 100 or more amino acid residues) are referred to as "proteins," and polymers having a small number of amino acid residues (e.g., less than 100 amino acids) are referred to as "peptides."

[0071] In this specification, the terms "polynucleotide" and "nucleic acid" are used interchangeably to refer to nucleotide polymers to which nucleotides are bound by phosphodiester bonds. "Polynucleotide" and "nucleic acid" can be DNA, RNA, or a combination of DNA and RNA. In addition, "polynucleotide" and "nucleic acid" can be polymers of natural nucleotides, polymers of natural nucleotides and non-natural nucleotides (nucleotides (such as thiophosphate backbones) in which at least one portion of an analog, base portion, sugar portion, or phosphate portion of a natural nucleotide is modified), or polymers of non-natural nucleotides. Unless otherwise specified, base sequences are recorded from the 5' side to the 3' side.

[0072] In this specification, the term "gene" refers to a polynucleotide containing at least one open reading frame (ORF) encoding a specific protein. A gene may contain both exons and introns.

[0073] In this specification, the term "operably linked" as used in relation to a polynucleotide means that a first base sequence and a second base sequence are positioned in sufficient proximity such that the first base sequence can affect the second base sequence or a region under the control of the second base sequence. For example, "operably linked to a promoter" means that the polynucleotide is linked so that it is expressed under the control of the promoter.

[0074] In this specification, "a promoter is capable of functioning" means that the promoter is capable of expressing a polynucleotide operably linked to the promoter in cells of a subject.

[0075] In this specification, the term "expressible state" means that the polynucleotide or gene is in a state where it can be transcribed in a cell into which the polynucleotide has been introduced.

[0076] In the present specification, the term "expression vector" refers to a vector comprising a target polypeptide and having a system for expressing the target polynucleotide in cells into which the vector has been introduced.

[0077] In this specification, the term "drug delivery composition" refers to a composition for delivering a drug to any site (organ, organ, tissue, disease site, etc.) in a living body.

[0078] In this specification, "drug carrier" means a carrier for drug delivery. The drug carrier can be either organic or inorganic. When the drug carrier is composed of an organic substance, the drug carrier can be a cell.

[0079] In this specification, "a drug is enclosed in a cell" means that the drug is present in the cell and / or in the cell membrane. When the drug is present in the cell, the drug may be present inside a cell organelle.

[0080] In this specification, the phrase "a drug is locally present in a bag-like membrane structure" means that the majority of the drug is present inside the bag-like membrane structure of the subject (inside the bag) or in the membrane forming the bag-like membrane structure (hereinafter referred to as the "bag-like membrane"). In the case where the drug is locally present in the bag-like membrane structure possessed by the cell, the entire drug enclosed in the cell does not need to be present inside the bag-like membrane structure or in the bag-like membrane; a portion of the drug may also be present outside the bag-like membrane structure. In the case where the drug is "locally present in the bag-like membrane structure," the proportion of the drug present in the bag-like structure may be, for example, 50% or more of the total amount of the drug enclosed in the cell, preferably 60% or more, more preferably 70% or more, and even more preferably 80% or more.

[0081] In this specification, "low molecular weight compound" means a compound having a molecular weight of about 2000 or less. However, peptides and nucleic acids having a molecular weight of 2000 or less are not included in "low molecular weight compound".

[0082] In this specification, the term "synthetic polymer compound" refers to a non-natural compound with a molecular weight of 2000 or more. "Non-natural compound" refers to a compound that does not exist in nature. Examples of synthetic polymer compounds include various synthetic polymers (polyolefins, polyesters, polyamides, polyethylene glycol, poly(2-oxazoline), etc.). Artificially chemically synthesized peptides, proteins, and nucleic acids are not included in the term "synthetic polymer compound."

[0083] In this specification, "exogenous substance" means a substance introduced from outside the cell or a substance produced in the cell based on a substance introduced from outside the cell. Specific examples of substances produced in the cell based on a substance introduced from outside the cell include: transcription products (mRNA) and translation products (proteins) of foreign genes in cells into which foreign genes have been introduced, and active metabolites of prodrugs (drugs that exhibit the desired efficacy) in cells into which prodrugs have been introduced. Exogenous substances are substances that are different from substances originally possessed by the cell (endogenous substances).

[0084] In this specification, the term "drug" means a substance that shows beneficial activity in an organism. The beneficial activity shown by a drug is not particularly limited, and includes physiological activity, pharmacological activity, biological activity, and chemical activity useful for diagnosis, etc. For example, the activity may include: the pharmacological activity of a well-known compound as an active ingredient of a pharmaceutical product, and the chemical activity or physiological activity of a diagnostic drug administered to the body for use. As the activity, for example, immune induction activity, immune enhancement activity, anticancer activity, signal transduction inhibitory activity, signal transduction promoting activity, metabolic antagonist activity, analgesic activity, anti-inflammatory activity, bactericidal activity, antiviral activity, antiallergic activity, enzyme inhibitory activity, angiographic effect, fluorescent activity, etc. can be cited, but are not limited to these activities. A drug may also be a compound (so-called prodrug) that releases a compound that shows beneficial activity in an organism.

[0085] In this specification, the term "pharmaceuticals" includes pharmaceuticals for medical use and drugs taken in a broad sense for the treatment, prevention, or improvement of health. "Pharmaceuticals" may be registered or unregistered, and may be for medical or non-medical use.

[0086] In this specification, the term "variant" refers to a cell strain in which a mutation has occurred naturally or artificially in the genome (including the nuclear genome, chloroplast genome, and mitochondrial genome, hereinafter the same) of the original cell strain. The artificial method for causing the genome mutation is not particularly limited. Examples of such artificial methods include ultraviolet irradiation, radiation irradiation, chemical treatment with nitrous acid, and other chemical treatments, gene transfer, genome editing, and other genetic engineering methods.

[0087] In this specification, a "variant of the YFU3 strain" refers to an algae strain that has a mutation in the YFU3 strain genome and has both a diploid and haploid cell morphology. A "variant of the HKN1 strain" refers to an algae strain that has a mutation in the HKN1 strain genome and has both a diploid and haploid cell morphology.

[0088] In this specification, a "related species" refers to, for example, a cell strain whose base sequence of the rbcL gene, 18S rRNA gene, or 16s RNA gene is 90% or greater identical to the base sequence of the aforementioned gene in the original species. If the species is algae, the gene to be compared is the rbcL gene or the 18S rRNA gene, preferably the rbcL gene. The base sequence identity between the rbcL gene of the original algae and the base sequence of the rbcL gene of the related algae is preferably 95% or greater, more preferably 97% or greater, even more preferably 98% or greater, and particularly preferably 99% or greater. The base sequence of the rbcL gene of the algae can be obtained using well-known methods. For example, DNA can be extracted from cells of the target algae using well-known methods, amplified using PCR or other methods, and the base sequence of the amplified DNA fragment can be analyzed using a DNA sequence analyzer to obtain the base sequence of the rbcL gene of the target algae.

[0089] [Drug delivery composition]

[0090] In one embodiment, the present invention provides a drug delivery composition comprising acid-resistant cells encapsulating a drug. In a preferred embodiment, the drug is locally present in the bag-like membrane structure of the acid-resistant cells.

[0091] <Acid-resistant cells>

[0092] In this specification, "acid-resistant cells" refer to cells resistant to acidic conditions. Specifically, the acidic conditions include pH conditions of 1 to 3. Acid-resistant cells are preferably resistant to pH conditions of 1 to 4, and more preferably resistant to pH conditions of 1 to 5.

[0093] "Resistant to acidic conditions" means that cells do not rupture and elution of cell contents occurs under acidic conditions.

[0094] Acid resistant cells can be living cells or dead cells, but are preferably cells that maintain the form of the cell. Acid resistant cells are preferably cells in which cell membrane and / or its outer membrane are not damaged, in which the dissolution of cell content does not occur. When acid resistant cells are living cells, the cells can grow under acidic conditions.

[0095] The type of acid-resistant cells is not particularly limited. Examples of acid-resistant cells include acid-resistant algal cells. Preferred examples of such algal cells include cells of microalgae isolated from acidic environments such as acidic hot springs. Specific examples of such microalgae include algae belonging to the class Cyanidiophyceae.

[0096] Hot spring red algae are taxonomically classified as Rhodophyta and Cyanidiophyceae. Within the hot spring red algae, there are currently three genera: Cyanidioschyzon, Cyanidium, and Galdieria. Acid-resistant cells can belong to any of these genera. For example, systematic analysis using base sequences of the 18S rRNA gene or the chloroplast ribulose 1,5-bisphosphate carboxylase / oxygenase large subunit (rbcL) gene can determine whether a particular alga belongs to the hot spring red algae. Systematic analysis can be performed using well-known methods. Figure 7 ] shows a molecular phylogenetic tree based on the base sequence of the rbcL gene of algae belonging to the hot spring red algae class.

[0097] Among algae belonging to the class Rhodophyta, some have both diploid and haploid cell forms. Diploid cells can produce haploid cells through meiosis. Furthermore, it is believed that haploid cells produce diploid cells through the fusion of two haploid cells.

[0098] The haploid cell is compared with the diploid cell, and it is easy to use the preparation of the transformant of genetic recombination technology.Therefore, as described below, when the gene of the peptide of coding as medicine is imported into the acid-resistant cell, can preferably use the haploid cell.In addition, use the haploid cell to prepare a plurality of transformants that have imported any drug encoding gene, by hybridizing between these transformants, can prepare thus the diploid that has a plurality of drug encoding genes concurrently, inner bag a plurality of medicines.

[0099] Determining whether algae are diploid or haploid can be done by confirming the copy number of the same locus. That is, if the copy number of the same locus is 1, it is determined to be haploid. Alternatively, algae can be determined to be haploid using a next-generation sequence analyzer or the like. For example, sequence reads of the entire genome are obtained using a next-generation sequence analyzer or the like, and after assembling these sequence reads, the sequence reads are mapped to the assembled sequence. In diploids, base differences between alleles can be found in various regions of the genome, but in haploids, since only a single allele exists, such regions cannot be found.

[0100] Alternatively, cells can be stained with a nuclear staining reagent such as DAPI and compared with cells known to be haploid. Cells showing equivalent fluorescence brightness can be determined as haploid, while cells showing approximately twice the fluorescence brightness can be determined as diploid. Alternatively, cells can be stained with a nuclear staining reagent such as DAPI and compared with cells known to be diploid. Cells showing equivalent fluorescence brightness can be determined as diploid, while cells showing approximately 1 / 2 the fluorescence brightness can be determined as haploid.

[0101] In order to release the drug quickly in the intestine, the acid-resistant cells preferably do not have a strong cell wall. In this specification, "do not have a strong cell wall" means that the cells are disrupted in any of the following cell disruption treatments (A) to (C).

[0102] (A) Cells were suspended in an isotonic solution with a pH of 7 or higher and left for more than one week.

[0103] (B) Cells were suspended in distilled water for more than 1 minute.

[0104] (C) The cells were dried and then suspended in an isotonic solution with a pH of 7 or higher.

[0105] In the above (A) to (C), when the cells are cultured cells, the culture medium may be removed by centrifugation or the like before each treatment, and the algal cells may be washed with an isotonic solution or the like.

[0106] In the above (A) and (C), examples of the isotonic solution include a buffer solution at pH 7 containing 10% sucrose and 20 mM HEPES.

[0107] In (C) above, examples of drying treatment include drying in a refrigerator (4°C) and freeze drying. The algal cell pellet recovered by centrifugation is used for the drying treatment. When drying in a refrigerator, the drying treatment time varies depending on the cell mass, but can be 3 days or longer.

[0108] Alternatively, the cell suspension after the cell disruption treatments (A) to (C) above is centrifuged (1500 × g, 3 minutes) and the ratio of the protein mass in the supernatant to the total protein mass in the cell suspension is calculated to determine whether cell disruption has occurred. Specifically, cell disruption is determined to have occurred when the disruption ratio calculated using the following formula is 20% or greater.

[0109]

[0110] Alternatively, cells in the cell suspension may be observed using an optical microscope (eg, 600x magnification) and cell disruption may be determined when the ratio of disrupted cells is approximately 10% or more, preferably approximately 20% or more, of the total cells.

[0111] If the cells do not have a strong cell wall, the cell wall is generally not visible under optical microscopic observation (e.g., 600x magnification). Furthermore, whether or not the cells are disrupted by mild hypotonic treatment at a pH of 6 or below does not affect the determination of whether the algae do not have a strong cell wall.

[0112] In the cell disruption treatments (A) and (C), an isotonic solution at a pH of 7 or higher can be used. Therefore, the cells disrupted in either of the cell disruption treatments (A) and (C) can be said to be cells that have been disrupted at a pH of 7 or higher. In order to rapidly release the drug in the intestine, the acid-resistant cells are preferably cells that have been disrupted at a pH of 7 or higher.

[0113] Alternatively, by immersing the cells in a buffer solution at pH 7 or higher and observing for about 10 to 30 minutes to confirm whether the algal cells are ruptured, it can be determined whether the cells are ruptured under conditions of pH 7 or higher.

[0114] Examples of acid-resistant cells with these characteristics include haploids of Cyanidioschyzon merolae and Galdieria, and haploids of Cyanidium, among algae belonging to the class Rhodophyceae. These algae can be isolated from acidic environments such as acidic hot springs, or obtained from culture collections. Examples of such collections include the National Institute for Environmental Studies, Microbial System Collection Facility (16-2, Onogawa, Tsukuba, Ibaraki, Japan), and the American Type Culture Collection (ATCC; 10801 University Boulevard, Manassas, VA 20110, USA).

[0115] Examples of haploid algae belonging to the genus Galdieria include haploids of Galdieria sulphuraria and Galdieria partita, as well as haploids of their closely related species, variants, and progeny. For example, diploid algae belonging to the genus Galdieria obtained from a culture collection, etc., can be cultured until reaching a stationary phase, and then continued for an arbitrary period of time to produce haploid cells in the culture medium. These haploid cells can be recovered and used as acid-resistant cells.

[0116] Examples of haploids of algae belonging to the genus Cyanidium include haploids of Cyanidium sp. YFU3 (FERM BP-22334) (hereinafter referred to as "YFU3") and haploids of Cyanidium sp. HKN1 (FERM BP-22333) (hereinafter referred to as "HKN1"), and their closely related species, mutants, and progeny.

[0117] Strain YFU3 (haploid) is a single-celled red algae isolated from the high-temperature, acidic waters of a hot spring in Yufu City, Oita Prefecture, Japan. Strain YFU3 was deposited with the Japan Patent Organisms Depository (KOKU), Japan (2-5-8, Kazusa Kamata, Kisarazu-shi, Chiba Prefecture, Japan) on May 30, 2017, under the accession number FERM P-22334. It was transferred to an international depository on April 20, 2018, under the accession number FERM BP-22334.

[0118] The HKN1 strain is a single-celled red alga isolated from the hot, acidic waters of a hot spring in Hakone-machi, Ashigarashimo-gun, Kanagawa Prefecture, Japan. The HKN1 strain (haploid) was deposited with the Japan Patent Organisms Depository (KPMD) on May 30, 2017, under the accession number FERM P-22333. It was transferred to the international depository on April 20, 2018, under the accession number FERM BP-22333.

[0119] Algae belonging to the hot spring red algae can be cultured using a culture medium for microalgae culture. The culture medium is not particularly limited, and examples thereof include inorganic salt culture media containing a nitrogen source, a phosphorus source, and trace elements (zinc, boron, cobalt, copper, manganese, molybdenum, iron, etc.). For example, as nitrogen sources, ammonium salts, nitrates, nitrites, urea, ammonia, etc. can be mentioned, and as phosphorus sources, phosphates, etc. can be mentioned. Examples of such culture media include 2×Allen medium (Allen MB. Arch. Microbiol. 1959 32: 270-277.), M-Allen medium (Minoda A et al. Plant Cell Physiol. 2004 45: 667-71.), and MA2 medium (Ohnuma M et al. Plant Cell Physiol. 2008 Jan; 49(1): 117-20.).

[0120] Algae belonging to the class Rhodophyta can also be cultured in a culture medium using acidic hot spring drainage. "Acidic hot spring drainage" refers to acidic drainage discharged from hot spring facilities. While the acidic hot spring drainage is not particularly limited, it is preferably at a pH of 1.0 to 4.0, more preferably 1.0 to 3.0. "Culture medium using acidic hot spring drainage" refers to a culture medium prepared by adding a nitrogen source, a phosphorus source, trace elements, and the like to the acidic hot spring drainage. Culture medium using acidic hot spring drainage is preferably a culture medium obtained by adding a nitrogen source to the acidic hot spring drainage, more preferably a culture medium obtained by adding a nitrogen source and a phosphorus source (for example, see Hirooka S and Miyagishima SY (2016) Cultivation of Acidophilic Algae Galdieria sulphuraria and Pseudochlorella sp. YKT1 in Media Derived from Acidic Hot Springs. Front Microbiol. Dec 20; 7: 2022.). As nitrogen sources, ammonium salts (ammonium sulfate, etc.), urea, nitrates (sodium nitrate, etc.) and the like can be cited, preferably ammonium salts and urea, more preferably ammonium salts. As the amount of nitrogen source added, for example, 1 to 50 mM can be cited as the amount of nitrogen added. As for the amount of nitrogen source added, 5 to 40 mM is preferred, more preferably 10 to 30 mM is preferred, as a nitrogen source. As for phosphorus sources, phosphates (potassium dihydrogen phosphate, etc.) can be cited. As for the amount of phosphorus source added, 0.1 to 10 mM can be cited as the amount of phosphorus added, and 0.5 to 5 mM is preferred, more preferably 1 to 3 mM is preferred, as a phosphorus added. As algae belonging to the hot spring red algae class, since they can also be cultured using a culture medium using acidic hot spring drainage, acidic hot spring drainage can be effectively utilized and can be cultured at a low cost.

[0121] When the algae belonging to the genus Galdieria are algae, the nitrogen source is preferably an ammonium salt or urea, and more preferably an ammonium salt. When the algae belonging to the genus Cyanidium are algae, the nitrogen source is preferably an ammonium salt or nitrate, and more preferably an ammonium salt.

[0122] As mentioned above, algae belonging to the hot spring red algae class can grow at high densities under relatively relaxed culture conditions. Examples of pH conditions include pH 1.0 to 6.0, preferably pH 1.0 to 5.0. When culturing outdoors, to prevent the proliferation of other organisms, it is preferable to cultivate under highly acidic conditions, such as pH 1.0 to 3.0.

[0123] Examples of temperature conditions include 15 to 50° C., preferably 30 to 50° C. When culture is performed outdoors, culture is preferably performed at a high temperature to prevent the proliferation of other organisms, and examples of such conditions include 35 to 50° C.

[0124] As the light intensity, 5 to 2000 μmol / m 2 s, preferably 5 to 1500 μmol / m 2 When growing outdoors, they can be grown in sunlight. When growing indoors, they can be grown under continuous light or with a light-dark cycle (e.g., 10L:14D).

[0125] <Medications>

[0126] The drug contained within the acid-resistant cells is not particularly limited and may be any drug. Examples of the drug include, but are not limited to, low molecular weight compounds, peptides, proteins, nucleic acids, lipids, carbohydrates, vitamins, hormones, and synthetic polymer compounds. Preferably, the drug is at least one drug selected from the group consisting of low molecular weight compounds, peptides, proteins, and nucleic acids.

[0127] As low molecular weight compounds, low molecular weight compounds known to the public as active ingredients of pharmaceuticals can be used without particular limitation. Low molecular weight compounds can be contrast agents, fluorescent pigments, etc. for diagnostic drugs. As low molecular weight compounds, for example, immunopotentiators, anticancer agents, signal transduction inhibitors, metabolic antagonists, analgesics, anti-inflammatory agents, antibiotics, antiallergic agents, central nervous system disease therapeutic drugs, circulatory organ disease therapeutic drugs, respiratory organ system disease therapeutic drugs, digestive organ system disease therapeutic drugs, urogenital organ disease therapeutic drugs, etc., contrast agents, fluorescent pigments, etc. can be cited, but are not limited to these. Low molecular weight compounds are not limited to active ingredients of pharmaceuticals, and can also be ingredients in food (such as nutrients such as amino acids and vitamins) and food additives (spices, etc.).

[0128] Examples of nucleic acids include nucleic acid molecules used as nucleic acid medicines (siRNA, miRNA, antisense RNA, aptamers, decoys, CpG oligonucleic acids, etc.).

[0129] Examples of synthetic polymers include industrially produced polymers such as polyolefins, polyesters, and polyamides, and polymers in granular or spherical forms. Among these, some are expected to have immunopotentiating effects.

[0130] The drug can be a microcapsule containing a low molecular weight compound, or a sustained-release microcapsule or a microcapsule that releases the drug depending on the environment such as temperature, pH, and pressure.

[0131] As peptides or proteins (hereinafter also collectively referred to as "drug peptides"), peptides and proteins known as active ingredients of pharmaceuticals can be used without particular limitation. Examples of drug peptides include, but are not limited to, antigens, cytokines, growth factors, hormones, enzymes, antibodies, antibody fragments, ligands, and blood component proteins.

[0132] Among these, pharmaceutical peptides are preferably immunogenic. "Immunogenicity" of a pharmaceutical peptide means that it can induce immunity against the pharmaceutical peptide in a living organism to which it is administered. The immunity induced by the pharmaceutical peptide can be cellular immunity, humoral immunity, or both.

[0133] The pharmaceutical peptide is more preferably a pharmaceutical peptide that contributes to intestinal immunity. "Intestinal immunity" refers to the biological defense system used to prevent foreign matter from invading the body from the intestine. The intestinal immune system is composed of lymphoid tissue such as Peyer's patches, immunocompetent cells in the lamina propria of the mucosa, intestinal epithelial cells, and lymphocytes present therein. The pharmaceutical peptide that contributes to intestinal immunity can be a pharmaceutical peptide that acts on any one or more of these intestinal immune systems and strengthens the intestinal immune system.

[0134] Examples of pharmaceutical peptides that contribute to intestinal immunity include immunogenic peptides or immunogenic proteins of pathogenic microorganisms or pathogenic viruses (hereinafter collectively referred to as "pathogens"). The immunogenic pharmaceutical peptide can be appropriately selected based on the infectious disease to which the drug delivery composition of this embodiment is intended. Immunogenic peptides or immunogenic proteins are also referred to as antigenic peptides or antigenic proteins.

[0135] For example, when the drug delivery composition of this embodiment is applied to humans, immunogenic peptides or immunogenic proteins of human pathogens can be used as drug peptides. Examples of human pathogens include, but are not limited to, rabies virus, rotavirus, influenza virus, HIV, poliovirus, hepatitis A virus, hepatitis B virus, human papillomavirus, Vibrio cholerae, Salmonella, tuberculosis, Streptococcus pneumoniae, anthrax, and Salmonella typhi.

[0136] For example, when the drug delivery composition of this embodiment is applied to livestock, immunogenic peptides or immunogenic proteins of livestock pathogens can be used as drug peptides. Examples of livestock pathogens include, but are not limited to, rabies virus, bovine rotavirus, bovine coronavirus, Akabane disease virus, bovine adenovirus, bovine parainfluenza virus, bovine Salmonella, tuberculosis, porcine circovirus, swine influenza virus, porcine parvovirus, swine cholera virus, and Streptococcus suis.

[0137] For example, immunogenic peptides or immunogenic proteins can be designed using full-length proteins or partial peptides of proteins constituting the outer membrane or capsid of pathogenic viruses or full-length proteins or partial peptides of cell membrane proteins of pathogenic bacteria. For example, when the pathogen is rabies virus, the full length or partial peptides of the glycoprotein (base sequence is SEQ ID NO: 1, amino acid sequence is SEQ ID NO: 2) can be exemplified as immunogenic proteins.

[0138] (Localization of drugs into pocket-like membrane structures)

[0139] In the cell of acid-resistant cell, preferred medicine is locally present in the bag-like membrane structure that acid-resistant cell has.In this manual, the meaning of " bag-like membrane structure " refers to the structure that is divided into bag-like by biomembrane or biomembrane simulation structure, as specific example, can enumerate cell membrane, organelle and exogenous liposome etc.As organelle, for example can enumerate mitochondria, chloroplast, endoplasmic reticulum, vacuole, nucleus, peroxisome and Golgi body etc., but are not limited to these." exogenous liposome " means the liposome that imports cell from the outside.

[0140] The drug's local presence in the pouch-like membrane structure of acid-resistant cells inhibits its degradation by cytoplasmic enzymes. Consequently, the drug is protected from degradation by intracellular enzymes until the acid-resistant cells are delivered to a specific site in the body (e.g., the intestine) and rupture.

[0141] There is no particular limitation on the method for making the drug locally present in the bag-like membrane structure. For example, a method can be cited that utilizes a signal peptide (hereinafter referred to as a "mobility signal") that indicates movement to any bag-like structure or a protein (hereinafter referred to as a "mobility protein") that moves to the bag-like structure. For example, a mobile signal or mobile protein targeting any bag-like structure is combined with a drug and introduced into acid-resistant cells, thereby enabling the drug to be locally present in the bag-like structure. For example, in the case of making the drug locally present in any part of mitochondria, vacuoles, peroxisomes, endoplasmic reticulum, cell membrane, Golgi apparatus and cell nucleus, the drug can be combined with a mobile signal (signal peptide) or mobile protein targeting mitochondria, vacuoles, peroxisomes, endoplasmic reticulum, cell membrane, Golgi apparatus or cell nucleus. For these mobile signals and mobile proteins, a variety of well-known mobile signals and mobile proteins can be selected according to the type of acid-resistant cells. Alternatively, the pocket membrane structure where the drug is desired to be localized can be separated from acid-resistant cells by cell fractionation methods such as density gradient centrifugation, and the proteins in the pocket membrane structure can be analyzed to obtain the transport signal or transport protein for the pocket membrane structure.

[0142] For example, when Cyanidioschyzon merolae is used as the acid-resistant cell, the following migration signals or migration proteins can be used as migration signals or migration proteins.

[0143] As a chloroplast-specific mobility protein, a protein composed of the N-terminal 130 residues (base sequence: sequence number 5, amino acid sequence: sequence number 6) of the chloroplast preprotein translocase SecA subunit (CMQ393C; base sequence: sequence number 3, amino acid sequence: sequence number 4) can be used (Sumiya et al 2016, Proc Natl Acad Sci US A. 113(47): E7629-E7638; PMID: 27837024).

[0144] As a migration signal for the mitochondrial matrix, a peptide consisting of the N-terminal 78 residues (base sequence: SEQ ID NO: 9, amino acid sequence: SEQ ID NO: 10) of EF-TU (CMS502C) (base sequence: SEQ ID NO: 7, amino acid sequence: SEQ ID NO: 8) can be used (Imoto et al 2013, BMJ. 300(6735): 1316-8; PMID: 2369666).

[0145] As a protein for vacuole movement, prenylated Rab receptor PRA1 (CMJ260C) (base sequence is sequence number 7, amino acid sequence is sequence number 8), ABC transporter (CMS401C) (base sequence is sequence number 13, amino acid sequence is sequence number 14) or O-methyltransferase (CMT369C) (base sequence is sequence number 15, amino acid sequence is sequence number 16) can be used (Yagisawa et al 2009, Plant J. 60 (5): 882-93; PMID: 19709388).

[0146] As a peroxisome-targeting protein, catalase (CMI050C) (base sequence: SEQ ID NO: 17, amino acid sequence: SEQ ID NO: 18) (Moriyama et al 2014, Planta. 240(3): 585-98; PMID: 25009310) can be used.

[0147] As endoplasmic reticulum-targeting mobility proteins, ACC1 (CMM188C) (base sequence is sequence number 19, amino acid sequence is sequence number 20), PAP (CMT239C) (base sequence is sequence number 21, amino acid sequence is sequence number 22) or ALA1 (CMR396C) (base sequence is sequence number 23, amino acid sequence is sequence number 24) can be used (Mori et al 2016, Front Plant Sci. 7: 958; PMID: 27446184).

[0148] ALA1 (CMR396C) can be used as a transport protein targeting the cell membrane (Mori et al 2016, Front Plant Sci. 7:958; PMID:27446184). Since ALA1 (CMR396C) is also a transport protein targeting the endoplasmic reticulum, using ALA1 (CMR396C) allows the drug to be localized to both the cell membrane and the endoplasmic reticulum.

[0149] As a Golgi apparatus-targeting protein, Got1 (CMI302C) (base sequence: SEQ ID NO: 25, amino acid sequence: SEQ ID NO: 286) and the like can be used (Yagisawa et al 2013, Protoplasma. 250(4): 943-8; PMID: 23197134).

[0150] As a protein that moves to the cell nucleus, topoisomerase I type IB (CMM263C) (base sequence: SEQ ID NO: 27, amino acid sequence: SEQ ID NO: 28) and the like can be used (Moriyama et al 2014, Genome Biol Evol. 6(1): 228-37; PMID: 24407855).

[0151] When the drug is a peptide drug, it can be internalized in acid-resistant cells as a fusion protein with a migration signal or migration protein. By fusion of the peptide drug with the migration signal or migration protein, the drug can be localized in the pocket-like membrane structure targeted by the migration signal or migration protein.

[0152] For example, by importing a gene encoding a fusion protein of a drug peptide and a mobile signal or a mobile protein (hereinafter also referred to as "fusion protein gene") into an acid-resistant cell and expressing the fusion protein in the acid-resistant cell, the fusion protein moves to the bag-like membrane structure with the mobile signal or the mobile protein as the target. As a result, the drug peptide contained in the fusion protein is locally present in the bag-like membrane structure. Therefore, in a preferred embodiment, the acid-resistant cell is a cell into which a fusion protein gene comprising a mobile signal or a mobile protein and a drug peptide has been imported in an expressible state, and is a cell having the fusion protein gene. In addition, in a preferred embodiment, the acid-resistant cell is a cell expressing the fusion protein gene.

[0153] In addition to the coding sequence for the drug peptide and the coding sequence for the migration signal or migration protein, the fusion protein gene may also include a sequence encoding a peptide that enhances intestinal cell recognition. Examples of peptides that enhance intestinal cell recognition include the Col peptide (SEQ ID NO: 43).

[0154] The fusion protein gene of drug peptide and mobile signal or mobile protein is preferably operably connected to the promoter that can play a role in acid-resistant cells. As long as the promoter is a promoter that can play a role in acid-resistant cells, there is no particular limitation. From the viewpoint of maintaining the amount of drug in the cell, the promoter of the housekeeping gene with a large expression amount is preferably used. For example, when acid-resistant cells are Cyanidioschyzon merolae, as a promoter, for example, the promoter (example, -600 to -1; "-1" represents the nucleotide immediately before the start codon) of APCC (CMO250C), the promoter of CPCC (CMP166C), the promoter of catalase (CMI050C) etc. can be suitable for use. The promoter sequence of APCC of Cyanidioschyzon merolae is shown in SEQ ID NO: 29, the promoter sequence of CPCC (CMP166C) of Cyanidioschyzon merolae is shown in SEQ ID NO: 30, and the promoter sequence of catalase (CMI050C) of Cyanidioschyzon merolae is shown in SEQ ID NO: 31. These promoters of Cyanidioschyzon merolae can also be used in other algae belonging to the class Hot Spring Rhodophyceae.

[0155] The gene encoding the fusion protein is introduced into acid-resistant cells in an expressible state, for example, in the form of an expression vector. In addition to the fusion protein and promoter, the expression vector may also include an enhancer, a poly A addition signal, a terminator, a 3'UTR, and other control sequences, as well as marker genes such as drug resistance genes. Examples of terminators and 3'UTRs include the 3'UTR of β-tubulin.

[0156] There is no particular limitation on the type of vector, and commonly used expression vectors can be appropriately selected and used according to the type of acid-resistant cells. The vector can be linear or circular, and can be a non-viral vector such as a plasmid, a viral vector (e.g., a retroviral vector such as a lentiviral vector), or a transposon-based vector.

[0157] When the acid-resistant cell is Cyanidioschyzon merolae, the URA5.3 gene (CMK046C) can be used as a selection marker. Among Cyanidioschyzon merolae, there is a Cyanidioschyzon merolae M4 strain (Minoda et al., Plant Cell Physiol. 2004 Jun; 45 (6): 667-71.), which is a mutant strain with uracil auxotrophy. The Cyanidioschyzon merolae M4 strain has a mutation in the URA5.3 gene and cannot synthesize uracil. Therefore, the Cyanidioschyzon merolae M4 strain cannot grow in a culture medium that does not contain uracil. Therefore, by using the Cyanidioschyzon merolae M4 strain as a parent strain and using the URA5.3 gene of the wild strain as a selection marker, it is possible to select a transformant into which the fusion gene has been introduced. More specifically, the fusion protein gene operably linked to a promoter is ligated to the URA5.3 gene set of a wild-type Cyanidioschyzon merolae strain (e.g., strain 10D) and introduced into the Cyanidioschyzon merolae M4 strain. Cells introduced with the fusion protein gene can then be obtained by culturing the cells in a medium containing no uracil.

[0158] There is no particular limitation on the method for introducing any fusion protein gene into acid-resistant cells, and well-known methods can be used. As gene introduction methods, for example, polyethylene glycol method, liposome transfection method, microinjection method, DEAE dextran method, gene gun method, electroporation method, calcium phosphate method etc. can be enumerated.

[0159] The fusion protein gene can be present as a plasmid in acid-resistant cells, or can be inserted into any of the nuclear genome, chloroplast genome, and mitochondrial genome. When the fusion protein gene is inserted into the genome, it can be inserted into a specific location of the genome or randomly inserted into the genome.

[0160] Homologous recombination can be used to insert a fusion protein gene into a specific genomic location. For example, the complete genome sequence of Cyanidioschyzon merolae has been deciphered (Matsuzaki M et al., Nature. 2004 Apr 8; 428(6983): 653-7), making it possible to insert the fusion protein gene into a desired location on the genome. The insertion location of the fusion protein gene in Cyanidioschyzon merolae is not particularly limited; for example, the region between CMD184C and CMD185C can be used.

[0161] In the fusion protein gene, the order of arranging the drug peptide and the mobilization signal or mobilization protein can be appropriately selected according to the type of the mobilization signal or mobilization protein. Generally, the coding sequence of the mobilization signal or mobilization protein is arranged on the 5' side of the coding sequence of the drug peptide.

[0162] When a gene encoding a drug peptide (hereinafter referred to as a "drug peptide gene") is inserted into the chloroplast genome or the mitochondrial genome, the drug peptide does not necessarily need to be a fusion protein with a migration signal or a migration protein. For example, the drug peptide gene is operably linked to a promoter that can function in the chloroplast, inserted into the chloroplast genome in an expressible state, and the drug peptide gene is expressed in the chloroplast, thereby enabling the drug peptide to be locally present in the chloroplast. Similarly, the drug peptide gene is operably linked to a promoter that can function in the mitochondria, inserted into the mitochondrial genome in an expressible state, and the drug peptide gene is expressed in the mitochondria, thereby enabling the drug peptide to be locally present in the mitochondria.

[0163] In the drug delivery composition of this embodiment, the drug is preferably localized in a cell organelle, more preferably in a chloroplast. Furthermore, the drug is preferably a drug peptide, preferably localized in a cell organelle targeted by the migration signal or migration protein in the form of a fusion protein with the migration signal or migration protein. The migration signal or migration protein is more preferably a chloroplast migration signal or a chloroplast migration protein.

[0164] <Optional ingredients>

[0165] The drug delivery composition of the present embodiment may also include other components in addition to comprising the acid-resistant cells. As other components, there are no particular limitations, for example, pharmaceutically acceptable carriers etc. can be cited. The meaning of "pharmaceutically acceptable carrier" refers to a carrier that does not hinder the function of the drug contained in the acid-resistant cells and does not show substantial toxicity to the subject being administered. The meaning of "does not show substantial toxicity" refers to that its components do not show toxicity to the subject being administered under the commonly used dosage. As pharmaceutically acceptable carriers, there are no particular limitations, and excipients, binding agents, disintegrants, lubricants, emulsifiers, stabilizers, diluents, oily bases, tackifiers, antioxidants, reducing agents, oxidants, chelating agents, solvents etc. can be cited. Pharmaceutically acceptable carriers can be used alone or in combination of two or more. Pharmaceutically acceptable carriers are preferably carriers that do not damage acid-resistant cells.

[0166] The drug delivery composition of the present embodiment can be suitably mixed with other components, is formed into the form of granules, tablets, jelly, liquid preparation, capsule etc. according to a universal method.In these dosage forms, it is preferably the dosage form that acid-resistant cells are not damaged, for example, it is preferably jelly, liquid preparation, capsule etc. For example, as shown in the embodiment described later, it is possible to use the form of the solidified body of the alginic acid comprising acid-resistant cells. In addition, except alginic acid, it is also possible to use the tackifiers or gelling agents such as gelatin, agar, carrageenan, locust bean gum, guar gum, xanthan gum, pectin, gellan gum, tamarind gum, gum arabic, the suspension comprising acid-resistant cells is solidified, as the drug delivery composition of the present embodiment. There is no particular limitation on the medium for the suspension of the acid-resistant cells, it is preferably the medium that does not cause cell rupture in the acid-resistant cells, preferably the isotonic solution of about pH 1~6. As the isotonic solution, for example, a culture medium for culturing acid-resistant cells and a glucose isotonic solution adjusted to about pH 1 to 6, a sucrose isotonic solution and various buffer solutions (phosphate buffered saline, HEPES buffer, citrate buffer, Tris buffer (tris(hydroxymethylaminomethane) buffer), etc.) can be cited. In one embodiment, the drug delivery composition is a solidified body of acid-resistant cells using a thickener or a gelling agent. By making the drug delivery composition a solidified body using a thickener and / or a gelling agent, the drying of acid-resistant cells can be prevented. "Solidified body of acid-resistant cells using a thickener or a gelling agent" refers to a solidified body obtained by gelling and solidifying a suspension of acid-resistant cells with a thickener or a gelling agent. In other words, "solidified body of acid-resistant cells using a thickener or a gelling agent" is a gel composition containing acid-resistant cells and at least one selected from the group consisting of a thickener and a gelling agent.

[0167] The route of administration of the drug delivery composition of this embodiment is not particularly limited and can be administered orally or parenterally, with oral administration being preferred. In the drug delivery composition of this embodiment, since the drug is encapsulated in acid-resistant cells, drug degradation by gastric acid can be inhibited. Therefore, the drug delivery composition of this embodiment is suitable for oral administration.

[0168] The drug delivery target of the drug delivery composition of the present embodiment is preferably intestinal (intestinal tract), more preferably small intestine. If the drug delivery composition of the present embodiment is orally administered, the drug is protected in the cell of acid-resistant cells and passes through the stomach. Then, when arriving at the intestine, due to the neutral to weakly alkaline pH conditions (more than pH 7) in the intestinal tract, cell rupture occurs in the acid-resistant cells, and the drug is released into the intestinal tract. The drug released into the intestinal tract works in the intestinal tract, contributes to the strengthening of intestinal immunity, etc. In addition, by the strengthening of intestinal immunity, it is also possible to expect to activate other mucosal immunity and systemic immunity.

[0169] As described above, the drug delivery composition according to this embodiment can suppress drug degradation in the stomach and deliver the drug to the intestine because the drug is encapsulated in acid-resistant cells. Furthermore, in acid-resistant cells, the drug is localized within the pouch-like membrane structure, protecting it from degradation by degradative enzymes in the cytoplasm.

[0170] Furthermore, by using acid-resistant cells that have been introduced with a drug peptide gene or a fusion protein gene containing the coding sequence for the drug peptide, it is possible to easily proliferate acid-resistant cells containing the drug. In particular, algae belonging to the class Rhodophyceae (hot spring red algae) can proliferate even in conditions of high acidity where other organisms cannot survive, and therefore can be cultivated outdoors in large quantities. Therefore, it is expected that manufacturing costs will be reduced.

[0171] [feed]

[0172] In one embodiment, the present invention provides a feed comprising the drug delivery composition of the above embodiment.

[0173] There are no particular limitations on the type of animals to which the feed of this embodiment can be administered, and examples include pets (dogs, cats, hamsters, rabbits, parrots, tropical fish, reptiles, amphibians, insects, etc.) and experimental animals (mice, rats, guinea pigs, etc.).

[0174] In addition to the drug delivery composition of the above embodiment, the feed of this embodiment may also contain other ingredients. As other ingredients, for example, commonly used feeds can be cited. For example, the drug delivery composition of the above embodiment can be added to existing feed as a feed additive. There is no particular limitation on the feed to which the drug delivery composition of the above embodiment is added, and it can be appropriately selected according to the target animal. By adding the drug delivery composition of the embodiment to a normal feed and giving it to an animal, the animal can ingest the drug through normal feeding behavior.

[0175] The drug delivery composition for the feed of the present embodiment can be any form, but in order to prevent the drug from leaking out from the acid-resistant cells, it is preferably a form that does not damage the cells of the acid-resistant cells. For example, the jelly, capsule, and the form solidified by gelling agent and / or tackifier etc. that have been exemplified above can be enumerated. When the drug delivery composition is added to feed as a feed additive, for example, as long as the solidified body of the drug delivery composition utilizing tackifier and / or gelling agent is adjusted to a suitable size and added to feed for mixing. Alternatively, after the drug delivery composition is added to feed and mixed, the mixture can be solidified using gelling agent and / or tackifier. The solidified body can be prepared into a suitable size as appropriate according to the size of the animal. By being formed into a solidified body utilizing tackifier and / or gelling agent, the drying of acid-resistant cells can be prevented.

[0176] There is no particular limitation on the content of the drug delivery composition of the above embodiment in the feed of the present embodiment, and it can be appropriately set according to the type of feed. For example, as the content of the drug delivery composition in the feed, 0.01 to 80 mass % can be exemplified, preferably 0.1 to 70 mass % can be exemplified, more preferably 0.1 to 60 mass % can be exemplified, and particularly preferably 0.1 to 50 mass % can be exemplified. As the content of acid-resistant cells in the feed, for example, 0.1 to 100 mg (wet weight) / g, 0.5 to 80 mg (wet weight) / g, 1 to 60 mg (wet weight) / g, etc. can be exemplified.

[0177] According to the feed of this embodiment, since it contains the drug delivery composition of the above embodiment, it is possible to ingest any drug as feed for animals. As described above, the drug delivery composition can protect any drug to avoid decomposition in the stomach and deliver it to the intestines. Therefore, by using a drug that works in the intestine in the drug delivery composition, the drug can be effectively made to act on the intestines of animals. In addition, when the drug is an immunogenic drug peptide, the intestinal immunity can be effectively activated for animals that have ingested the drug delivery composition. In addition, by activating the intestinal immunity, it is also possible to expect to activate other mucosal immunity and systemic immunity.

[0178] In another embodiment, the present invention provides a method for raising an animal, comprising feeding the animal a feed comprising the drug delivery composition according to the above embodiment.

[0179] Furthermore, in another embodiment, the present invention provides a method for imparting intestinal immunity to an animal, comprising allowing the animal to ingest a feed containing the drug delivery composition according to the above embodiment.

[0180] [Pharmaceuticals]

[0181] In one embodiment, the present invention provides a pharmaceutical product comprising the drug delivery composition of the above embodiment.

[0182] The pharmaceutical product of this embodiment can be a pharmaceutical product for humans or an animal. In the case of an animal pharmaceutical product, there is no particular limitation on the type of animal to which it can be applied. Examples include, but are not limited to, livestock (cattle, pigs, chickens, horses, sheep, goats, etc.), pets (dogs, cats, hamsters, rabbits, parrots, tropical fish, reptiles, amphibians, insects, etc.), aquatic animals (fish, shellfish, etc.), and experimental animals (mice, rats, guinea pigs, etc.).

[0183] The pharmaceutical product of this embodiment may further comprise other ingredients in addition to the drug delivery composition of the above embodiment. As other ingredients, there are no particular limitations, and pharmaceutically acceptable carriers may be cited. “Pharmaceutically acceptable carrier” means a carrier that does not interfere with the function of the drug and does not show substantial toxicity to the subject to which it is administered. In addition, “does not show substantial toxicity” means that its components do not show toxicity to the subject to which it is administered within the commonly used dosage. As pharmaceutically acceptable carriers, there are no particular limitations, and excipients, binders, disintegrants, lubricants, emulsifiers, stabilizers, diluents, oily bases, thickeners, antioxidants, reducing agents, oxidants, chelating agents, solvents, etc. may be cited. Pharmaceutically acceptable carriers may be used alone or in combination of two or more. Other ingredients may be ingredients other than those mentioned above, for example, pharmaceutical additives commonly used in pharmaceuticals may be used without particular limitations. In addition, other ingredients may also be active ingredients other than the drugs contained in the above-mentioned drug delivery composition. The active substance is not particularly limited, and examples thereof include intestinal regulating agents, anti-inflammatory agents, antibiotics, antibacterial substances, herbal medicines, structural promoters, antipyretics, analgesics, and the like.

[0184] The dosage form of the pharmaceutical product of this embodiment is not particularly limited, but to prevent drug leakage from the acid-resistant cells, it is preferably in a form that does not cause cell damage to the acid-resistant cells. Examples include tablets, granules, jelly, capsules, liquid preparations, and syrups. For example, the pharmaceutical product of this embodiment may also include a solidified form of the acid-resistant cells using a thickening agent and / or a gelling agent.

[0185] The content of the drug delivery composition of the above embodiment in the pharmaceuticals of the present embodiment is not particularly limited, and the content can be appropriately set according to the type of medicament contained in the drug delivery composition. For example, as the content of the drug delivery composition in pharmaceuticals, 0.01 to 80 mass % can be exemplified, preferably 0.1 to 70 mass % can be exemplified, more preferably 0.1 to 60 mass % can be exemplified, and particularly preferably 0.1 to 50 mass % can be exemplified. As the content of acid-resistant cells in pharmaceuticals, for example, 0.1 to 100 mg (wet weight) / g, 0.5 to 80 mg (wet weight) / g, 1 to 60 mg (wet weight) / g, etc. can be exemplified.

[0186] The administration route of the pharmaceutical of this embodiment is not particularly limited and can be oral or parenteral administration, preferably oral administration. Since the pharmaceutical of this embodiment is encapsulated in acid-resistant cells, it can inhibit the drug from being degraded by gastric acid.

[0187] The drug delivery target of the pharmaceutical of this embodiment is preferably the intestine (intestinal tract), more preferably the small intestine.

[0188] The pharmaceutical product according to the present embodiment, since it includes the drug delivery composition of the above embodiment, can protect any drug to avoid decomposition in the stomach, thereby enabling the drug to be delivered to the intestine. Therefore, by using a drug that works in the intestine for the drug delivery composition, the drug can be effectively made to act on the intestine. In addition, when the drug is an immunogenic drug peptide, it is possible to effectively activate intestinal immunity for animals that have ingested the drug delivery composition. In addition, by activating intestinal immunity, it is also possible to expect activation of other mucosal immunity and systemic immunity.

[0189] Therefore, the pharmaceutical product of this embodiment can be used for preventing and treating human diseases and improving health. It is particularly suitable for the following drugs: drugs that are preferably absorbed in the intestine rather than in the stomach, drugs that are decomposed or become insoluble by gastric acid, thereby hindering intestinal absorption, and drugs used for simultaneous intestinal absorption of multiple drugs.

[0190] In another embodiment, the present invention provides a method for administering a drug, comprising orally administering a pharmaceutical comprising the drug delivery composition according to the above embodiment to a subject.

[0191] Furthermore, in another embodiment, the present invention provides a method for imparting intestinal immunity to a subject, comprising orally administering to the subject a pharmaceutical comprising the drug delivery composition of the above embodiment.

[0192] [Drug carrier]

[0193] In one embodiment, the present invention provides a drug carrier comprising acid-resistant cells.

[0194] The acid-resistant cells contained in the drug carrier of this embodiment are the same as the acid-resistant cells described in the "<Acid-resistant cells>" of the above-mentioned "[Drug delivery composition]", and the same examples can be cited as preferred examples. The acid-resistant cells are resistant to acid and are not damaged even in an acidic environment such as the stomach. Therefore, by encapsulating the drug in the cells, it can be used as an acid-resistant drug carrier. The method for encapsulating the drug in the cells can be the same as the method described in the above-mentioned "[Drug delivery composition]". The drug carrier of this embodiment is preferably composed of acid-resistant cells.

[0195] The drug carrier of this embodiment can be suitably used for delivering drugs into the intestine, and can be suitably used for orally administered pharmaceuticals or orally ingested feeds.

[0196] [Medication capsules]

[0197] In one embodiment, the present invention provides a drug capsule containing a drug in the drug carrier of the embodiment.

[0198] The acid-resistant cells encapsulate drugs within their cells. As shown in the examples described below, drug release is virtually nonexistent in the acidic environment of the stomach. Therefore, the drug carrier containing the acid-resistant cells can be used as an acid-resistant drug capsule by encapsulating the drugs within the acid-resistant cells. The drug capsule of this embodiment is intended for drug delivery to the intestine and can be used as an oral drug capsule.

[0199] [Acid-resistant cells]

[0200] In one embodiment, the present invention provides an acid-resistant cell containing a drug. In a preferred embodiment, the drug is locally present in a bag-like membrane structure of the acid-resistant cell.

[0201] The acid-resistant cells of this embodiment are the same as those included in the drug delivery composition of the above embodiment, and preferred examples thereof may also be the same. Alternatively, the drug is locally present outside the bag-like membrane structure of the acid-resistant cells. In the case where the drug is locally present outside the bag-like membrane structure, the drug is present in the cytoplasm of the acid-resistant cells.

[0202] The drug is not particularly limited, and for example, it is preferably at least one drug selected from the group consisting of low molecular weight compounds, peptides, proteins and nucleic acids. For example, in the case of a drug that is affected by decomposition enzymes in the cytoplasm, it is preferred that the drug be locally present in the bag-like membrane structure. By being locally present in the bag-like membrane structure, the drug can be protected to avoid the influence of decomposition enzymes in the cytoplasm. Therefore, the drug can be effectively delivered to a specified part in the organism. For example, in the case of a drug that is a peptide, protein or nucleic acid, it is easily affected by proteases or nucleases in the cytoplasm; therefore, it is preferably locally present in the bag-like membrane structure. On the other hand, in the case of a drug (such as a low molecular weight compound) that is not easily affected by decomposition enzymes in the cytoplasm, the drug may also be locally present outside the bag-like membrane structure.

[0203] In one embodiment, the present invention provides an acid-resistant cell comprising an exogenous substance.

[0204] The acid-resistant cells of this embodiment are the same as the acid-resistant cells described in "<Acid-resistant cells>" of the above-mentioned "[Drug delivery composition]", and preferred examples are also the same.

[0205] There are no particular limitations on exogenous substances, and examples include, but are not limited to, drugs, poisons, dyes, fragrances, and compounds whose effects on organisms are unknown. There are no particular limitations on the method for introducing the exogenous substance into acid-resistant cells, and examples include methods of combining the exogenous substance with a cell-permeable substance (e.g., a cell-permeable peptide), and methods of encapsulating the exogenous substance within a cell-permeable micelle. Furthermore, when the exogenous substance is a drug, the same methods as those described in the "[Drug Delivery Composition]" above may be used.

[0206] The acid-resistant cells of this embodiment can be used, for example, to deliver exogenous substances. More specifically, the acid-resistant cells of this embodiment can be used in an oral composition for delivering exogenous substances into the intestine.

[0207] Furthermore, in another embodiment, the present invention provides a feed containing the acid-resistant cells.

[0208] Furthermore, in another embodiment, the present invention provides a pharmaceutical comprising the acid-resistant cells.

[0209] Furthermore, in another embodiment, the present invention provides a method for administering the exogenous substance, comprising orally administering the acid-resistant cells to a subject.

[0210] Furthermore, in another embodiment, the present invention provides a method for raising an animal, comprising allowing the animal to ingest the acid-resistant cells.

[0211] Furthermore, in another embodiment, the present invention provides a method for imparting intestinal immunity, comprising orally administering the acid-resistant cells.

[0212] [Method for producing acid-resistant cells]

[0213] In one embodiment, the present invention provides a method for producing acid-resistant cells containing a drug, comprising the step of introducing a gene encoding a fusion protein into the acid-resistant cells, wherein the fusion protein comprises: a peptide or protein serving as the drug, and a peptide or protein localized relative to the cell membrane or organelle.

[0214] The production method of this embodiment can be carried out by the method described in the above-mentioned "[Drug Delivery Composition] <Acid-Resistant Cells> (Localization of Drug to Bag-Like Membrane Structure)".

[0215] Example

[0216] Hereinafter, the present invention will be described with reference to examples, but the present invention is not limited to the following examples.

[0217] [Example 1]

[0218] (Preparation of GAPDH-GP-sfGFP expression strain)

[0219] In order to insert the DNA fragment of GAPDH-GP-sfGFP downstream of CMD184C (gene number) on the chromosome of Cyanidioschyzon merolae 10D, first, plasmid pD184-HSp-GAPDH-GP-sfGFP was prepared as shown below.

[0220] This plasmid was designed so that the following sequences were sequentially arranged in the multicloning site of the pQE80 plasmid (for maintenance and replication in Escherichia coli; manufactured by QIAGEN). The sequences were arranged in order from the 5' side as follows: the latter half of the CMD184C gene (773 bp-2773 bp of the gene open reading frame (ORF) and 25 bp downstream including the stop codon), the heat shock (HS) promoter (200 bp of the sequence upstream of the start codon of the HSP20 / CMJ101C gene; Sumiya et al 2014, Plos One. 22; 9(10): e111261; PMID: 25337786), GAPDH (1 bp-1209 bp of the open reading frame of the CMJ042C gene; GAPDH was described in Moriyama et al 2014, Planta. 240(3): 585-98; PMID: 25009310), and the rabies virus glycoprotein gene GP (ORF full length 1-1572 bp, UniProtKB accession No. P19462), the β-tubulin terminator (200 bp downstream of the stop codon of the β-tubulin / CMN263C gene), the URA selection marker, and the downstream of the CMD185 gene (the base sequence from 28 bp downstream of the stop codon to 1880 bp). The HS promoter is required to induce expression of GAPDH-GP-sfGFP by heating the culture medium. The URA selection marker is required for screening GAPDH-GP-sfGFP strains. In order to insert the DNA fragment downstream of CMD184C by homologous recombination, the second half and downstream sequences of CMD184C and the downstream of the CMD185C gene are required.

[0221] First, in order to prepare the plasmid pD184-HSp-GAPDH-GP-sfGFP, the following DNA fragments (1), (2), (3), (4), and (5) were prepared.

[0222] (1) Plasmid pD184-APCCp-EGFP-URA Cm-Cm(Containing pQE80 (SEQ ID NO: 32), the latter half of CMD184C (SEQ ID NO: 33), APCC promoter (SEQ ID NO: 34), EGFP (SEQ ID NO: 35), β-tubulin terminator (SEQ ID NO: 36), URA selection marker (SEQ ID NO: 37), and a DNA sequence downstream of the CMD185C gene (SEQ ID NO: 38); Fujiwara et al 2013, PLoS One. 8(9): e73608; PMID: 24039997) as a template, the primer set [#1d184(+25)R / #2bT3'(+1)F] was used to amplify the DNA sequence excluding the APCC promoter and EGFP by PCR. The base sequence of the DNA fragment (1) is shown in SEQ ID NO: 31.

[0223] (2) Using genomic DNA from C. merolae 10D as a template and a primer set [#3HS(-200)Fd184 / #4HS(-1)R], the DNA sequence of the HS promoter (SEQ ID NO: 39) was amplified by PCR.

[0224] (3) The GAPDH gene open reading frame (SEQ ID NO: 40) was amplified by PCR using the genomic DNA of C. merolae 10D as a template and the primer set [#5J042(1)Fhs / #6J042(1209)R-link3].

[0225] (4) The DNA sequence of GP was chemically synthesized (SEQ ID NO. 41) based on the codon usage frequency of C. merolae and amplified by PCR using the primer set [#7GP(1)F-linker3 / #8GP(1572)R-linker2] as a template.

[0226] (5) Using pAPCC-promoter-sfGFP-pmE2F-URA (Miyagishima et al 2014, Nat Commun. 5: 3807; PMID: 24806410) as a template and the primer set [#9sfGFP(1)F-linker2 / #10sfGFP(714)Rbt], sfGFP (SEQ ID NO: 42) was amplified by PCR.

[0227] The DNA fragments of (1), (2), (3), (4) and (5) were mixed and fused using the In-Fusion (registered trademark) HD ​​Cloning Kit (product number 639648, TAKARA) to replace pD184-APCCp-EGFP-URA Cm-GsThe HS promoter, GAPDH, GP and sfGFP were inserted into the APCC promoter and part of EGFP. After the InFusion reaction, the competent E. coli cells were introduced and the plasmid was amplified to obtain pD184-HSp-GAPDH-GP-sfGFP. Then, it was used as a template and the primer set [#11D184(1200)F / #12D184(+1400)R] was used to amplify the DNA fragment connected to the second half of the CMD184 gene (1200bp-2773bp of the gene ORF and 25bp downstream of the stop codon), HS promoter, GAPDH, GP, sfGFP, β-tubulin terminator, URA screening marker and the downstream of the CMD184C gene (base sequence from 28bp downstream of the stop codon to 1440bp) by PCR.

[0228] The DNA fragment was introduced into the uracil-auxotrophic strain M4 of C. merolae (Minoda et al 2004, Plant Cell Physiol. 45(6): 667-71.; PMID: 15215501) by the PEG method (Ohnuma et al 2008, Plant Cell Physiol. 49(1): 117-20; PMID: 18003671), and screening was performed using MA2 solid medium without uracil to obtain a GAPDH-GP-sfGFP-expressing strain.

[0229] (Evaluation of proteasome-induced degradation of GAPDH-GP-sfGFP protein)

[0230] The GAPDH-GP-sfGFP expressing strain of C. merolae prepared as described above (hereinafter referred to as “GAPDH-GP-sfGFP expressing strain”) was subcultured in 60 mL of MA2 medium in a conical flask at a cell density of OD750=0.2 and cultured under light irradiation (50 μmol m -2 s -1), and rotary culture was performed at 40°C for 2 days (before expression). Then, the culture solution was transferred to two Erlenmeyer flasks at a time of 20 mL. In order to induce GAPDH-GP-sfGFP gene expression by heat stimulation, the two Erlenmeyer flasks were transferred to a 50°C incubator and rotary culture was performed under light irradiation for 1 hour. Just before the transfer to 50°C, in order to inhibit the degradation of proteins by proteasomes, the proteasome inhibitor MG-132 (MG-132(+)) (Nishida et al 2005; Mol Biol Cell. 16(5): 2493-502; PMID: 15772156) was added to one of the two Erlenmeyer flasks at a final concentration of 100 μM. As a control, only 40 μL of DMSO, a solvent for MG-132, was added to the other Erlenmeyer flask (MG-132(-)). The expression of GAPDH-GP-sfGFP protein was confirmed by immunoblotting, and the effect of proteasome inhibition was verified by comparing the banding patterns. Anti-GFP antibody (clone JL-8, product number 632381, Takara) was used for the detection of GAPDH-GP-sfGFP protein.

[0231] Figure 1 The immunoblotting results are shown in Figure 1. In MG-132(-), the GAPDH-GP-sfGFP protein band is thinner than in MG-132(+). This result indicates that the GAPDH-GP-sfGFP protein is partially degraded by the proteasome after expression.

[0232] (Analysis of Intracellular Localization of GAPDH-GP-sfGFP Protein)

[0233] To analyze the intracellular localization of the GAPDH-GP-sfGFP protein, the GAPDH-GP-sfGFP expression strain was cultured at 50°C in the presence of MG-132 under light irradiation for 1 hour, and the fluorescence of the GAPDH-GP-sfGFP protein was observed using a fluorescence microscope.

[0234] Figure 2 A fluorescence microscope image of a GAPDH-GP-sfGFP expression strain is shown in Figure 1. The fluorescence signal of sfGFP indicates that the GAPDH-GP-sfGFP protein is localized in the cytoplasm. Figure 2 Image (PC) in (A) is a phase contrast microscope image showing the outline of cells. Figure 2 The image (Chl) in (B) is the autofluorescence image of chloroplasts. Figure 2 Image (C) (sfGFP) is a fluorescence image of sfGFP.

[0235] [Example 2]

[0236] (Preparation of Chl-TP-3HA-GP-Col Expression Strain)

[0237] In order to express Chl-TP-3HA-GP-Col (refer to Figure 3 ) was inserted downstream of CMD184C (gene number) on the chromosome of C. merolae 10D, and first, plasmid pD184-APCCp-Chl-TP-3HA-GP-Col was prepared as shown below.

[0238] This plasmid was designed so that the following sequences were arranged in order from the 5' side in the multiple cloning site of the pQE80 plasmid. The sequences are arranged in order from the 5' side as follows: the second half of the CMD184C gene (773 bp-2773 bp of the gene ORF and 25 bp downstream including the stop codon), the APCC promoter (600 bp of the sequence upstream of the start codon of the APCC / CMO250C gene), the chloroplast movement signal Chl-TP (1 bp-390 bp of the SECA / CMQ393C gene ORF; Sumiya et al 2016, Proc Natl Acad Sci USA. 113(47):E7629-E7638; PMID: 27837024), a sequence encoding a 3xHA tag (for confirming expression using an HA antibody), a rabies virus glycoprotein gene GP (1572 bp, UniProtKB accession No. P19462), a sequence encoding a Col peptide (Col peptide: SFHQLPARSPLP (SEQ ID NO: 43), a peptide that enhances antigen recognition by M cells involved in intestinal immunity; Kim et al 2010, J Immunol. 185(10): 5787-95; PMID: 20952686), β-tubulin gene terminator (200 bp downstream of the stop codon of β-tubulin / CMN263C gene), URA Cm-Gs Screening marker and the downstream of CMD185 gene (base sequence from 28bp to 880bp downstream of the stop codon). In order to insert the DNA fragment into the downstream of CMD184C by homologous recombination, the latter half and downstream sequence of CMD184C and the downstream of CMD185 gene are necessary. In order to continuously express Chl-TP-HA-GP-Col, APCC promoter is necessary (Watanabe et al 2011, J Gen Appl Microbiol. 57(1): 69-72; PMID: 21478650). In order to screen transformants with Chl-TP-3HA-GP-Col inserted, URACm-Gs A selection marker is essential (Imamura et al 2010, Plant Cell Physiol. 51(5):707-17; PMID: 20375110), and protein expression can be increased by making the gene multi-copy (Fujiwara et al 2013, PloS One 8(9):e73608; PMID: 24039997).

[0239] In order to prepare the plasmid pD184-APCCp-Chl-TP-HA-GP-Col, the following DNA fragments (1), (2), (3) and (4) were prepared.

[0240] (1) Plasmid pD184-APCCp-EGFP-URA Cm-Gs (including pQE80 (SEQ ID NO: 32), the second half of CMD184C (SEQ ID NO: 33), APCC promoter (SEQ ID NO: 34), EGFP (SEQ ID NO: 35), β-tubulin terminator (SEQ ID NO: 36), URA Cm-Gs The screening marker (SEQ ID NO: 44) and the DNA sequence downstream of the CMD185 gene (SEQ ID NO: 38; Fujiwara et al 2013, PLoS One. 8(9): e73608; PMID: 24039997) were used as templates, and the primers [#13APCC(-1)R / #14bT3'(+1)] were used to amplify the DNA sequence excluding EGFP by PCR.

[0241] (2) The Chl-TP DNA sequence (SEQ ID NO: 45) was amplified by PCR using the genomic DNA of C. merolae 10D as a template and the primer set [#15SecA(1)Fapcc / #16SecA(390)R-linker-ha].

[0242] (3) Plasmid DNA containing 3xHA (SEQ ID NO: 46): pBSb-THA (Ohnuma et al 2008, Plant Cell Physiol. 49(1): 117-20; PMID: 18003671) was used as a template and primer set [#17HA(1)F / #18HA(90)R] was used to amplify 3xHA (SEQ ID NO: 46) by PCR.

[0243] (4) The ORF of GP was chemically synthesized (SEQ ID NO: 40) based on the codon usage frequency of C. merolae and amplified by PCR using the primer set [#19GP(1)Fha / #20Col-GP(1680)Rbt] as a template.

[0244] The DNA fragments of (1), (2), (3) and (4) were mixed and fused using the In-Fusion (registered trademark) HD ​​Cloning Kit (product number 639648, TAKARA) to replace pD184-APCCp-EGFP-URA Cm-Gs Chl-TP, 3x HA and rabies virus glycoprotein ORF were inserted into the EGFP part. After the InFusion reaction, the plasmid was introduced into E. coli competent cells and amplified to obtain pD184-APCCp-Chl-TP-3HA-GP-bt-URA Cm-Gs Then, using it as a template and primers [#11D184(1200)F / #12D 184(+1400)R], a DNA fragment connected to the latter half of the CMD184C gene (1200bp-2773bp of the gene ORF and 25bp downstream including the stop codon), APCC promoter, Chl-TP, 3xHA, GP, Col peptide, β-tubulin terminator, URACm-Gs selection marker and the downstream of the CMD185C gene (base sequence from base 28 to base 1440) was amplified by PCR.

[0245] The DNA fragment was introduced into the uracil-auxotrophic strain M4 of C. merolae (Minoda et al 2004, Plant Cell Physiol. 45 (6): 667-71; PMID: 15215501) by the PEG method (Ohnuma et al 2008, Plant Cell Physiol. 49 (1): 117-20; PMID: 18003671), and the strain was screened using MA2 solid culture medium that does not contain uracil to obtain the Chl-TP-3HA-GP-Col expression strain.

[0246] (Evaluation of proteasome-induced degradation of Chl-TP-3HA-GP-Col protein)

[0247] The ChlTP-sfGFP-HA-GP-Col expressing strain of C. merolae prepared as described above (hereinafter referred to as "Chl-TP-3HA-GP-Col expressing strain") and the wild-type strain (WT) as a negative control were subcultured in 60 mL of MA2 medium placed in an Erlenmeyer flask at a cell concentration of OD750 = 0.2 and irradiated with light (50 μmol m -2 s -1 ), and rotary culture was performed at 40°C for 2 days. Then, 20 mL of the culture medium of each strain was transferred to two conical flasks. In order to inhibit the protein degradation caused by the proteasome, the proteasome inhibitor MG-132 (MG-132(+)) (Nishida et al 2005; Mol Biol Cell.16(5):2493-502; PMID:15772156) was added to one of the two conical flasks at a final concentration of 100 μM. As a control, only 40 μL of DMSO (MG-132(-)), which is a solvent for MG-132, was added to the other conical flask. The expression of ChlTP-sfGFP-HA-GP-Col protein was confirmed by immunoblotting, and the effect of proteasome inhibition was verified by comparing the band patterns. Anti-HA antibody (clone 16B12, product number 901503, Biolegend) was used for the detection of ChlTP-sfGFP-HA-GP-Col protein.

[0248] Figure 4 The immunoblotting results are shown in . No difference in the banding pattern of the ChlTP-sfGFP-HA-GP-Col protein was observed between MG-132(-) and MG-132(+). This result indicates that the ChlTP-sfGFP-HA-GP-Col protein is not degraded by the proteasome.

[0249] (Analysis of Intracellular Localization of ChlTP-sfGFP-HA-GP-Co1 Protein)

[0250] To analyze the intracellular localization of the ChlTP-sfGFP-HA-GP-Col protein, the Chl-TP-3HA-GP-Col expression strain cultured for 2 days at 40°C under light irradiation in the absence of MG-132 was fixed and immunofluorescence staining was performed using an anti-HA antibody.

[0251] Figure 5 The results of immunofluorescence staining are shown in . The signal from the anti-HA antibody showed that the ChlTP-sfGFP-HA-GP-Co1 protein was localized in the chloroplast (between the thylakoid and the membrane in the central part). Figure 5 Image (PC) in (A) is a phase contrast microscope image showing the outline of cells. Figure 5 The image (Chl) in (B) is the autofluorescence image of chloroplasts. Figure 5 Image (C) (anti-HA) is an immunofluorescence staining image using an anti-HA antibody. It can be confirmed that the ChlTP-sfGFP-HA-GP-Col protein detected by the anti-HA antibody is localized in the chloroplast.

[0252] Table 1 shows the sequences of primers used in Examples 1 and 2.

[0253] [Table 1]

[0254]

[0255] [Example 3]

[0256] (GAPDH-GP-sfGFP expression strain administered to mice)

[0257] To become 1.3×10 8 The GAPDH-GP-sfGFP expression strain was suspended in 300mM glucose solution (isotonic solution) at a concentration of 1 cell / mL (OD750 = 4). 250μL of the suspension was delivered directly into the stomach of a mouse (ICR strain) using a probe. Subsequently, at 0, 0.5, and 1.0 hours, the stomach, upper small intestine, and lower small intestine were removed, and each organ was suspended in 1mL of 300mM glucose solution. After centrifugation of the suspension, the supernatant was collected and an enzyme-linked immunosorbent assay (ELISA assay) for sfGFP was performed, and the absorbance at 450nm was measured.

[0258] Table 2 shows the results of relative sfGFP concentration measurements (absorbance at 450 nm by enzyme-linked immunosorbent assay) in various organs. sfGFP was barely detectable in the stomach, but was detected in the small intestine immediately after administration. This result suggests that algal cells migrated from the stomach to the intestine immediately after administration, rupturing in the intestine without rupturing in the stomach.

[0259] [Table 2]

[0260]

[0261] [Example 4]

[0262] (Mice were fed alginate-solidified feed containing the sfGFP expression strain)

[0263] C. merolae 10D (Sumiya et al 2014, PLoS One. 9(10): e111261; PMID: 25337786) cells (sfGFP-expressing strain) in which sfGFP was expressed and labeled in the cytoplasm were mixed with a commercially available diet (CLEA Rodent Diet CE-2, CLEA Japan, Inc.), and the mixture was solidified with alginic acid as shown below to prepare a diet sample.

[0264] 27 mL of 300 mM glucose solution (isotonic solution) (OD750 = 4) in which the sfGFP expression strain was suspended was centrifuged at 3000 g for 10 minutes, and the precipitated cells were collected. The cells of the sfGFP expression strain and 1.12 g of commercial feed (CE-2) were suspended in 10 mL of a 2.5% sucrose solution containing 1% sodium alginate. Then, the suspension was dropped into a 10% calcium chloride solution to obtain a feed sample containing the sfGFP expression strain and an alginate solid body of the commercial feed. The content of the sfGFP expression strain in the feed sample was 4.6 mg wet weight / g (80-110 mg per grain).

[0265] Mice (ICR line) were allowed to freely ingest the feed sample for 4 hours and then raised normally. The gastrointestinal tract, upper small intestine and lower small intestine of the mice were removed 4, 8, 24 and 48 hours after the start of intake. The removed organs were suspended in a 300mM glucose solution, centrifuged at 1000g, and the supernatant was recovered as a sample for determining the extracellular concentration of sfGFP. After the supernatant was recovered, the precipitate was suspended again by adding distilled water (DW) equal to the recovered supernatant, and after centrifugation at 1000g, the supernatant was recovered as a sample for determining the intracellular concentration of sfGFP. The amount of sfGFP in the extracellular concentration determination sample and the intracellular concentration determination sample was quantified using a commercially available ELISA kit (GFP ELISA kit; cat no.ab171581, abcam) as the extracellular concentration and intracellular concentration, respectively.

[0266] Table 3 shows the results of relative sfGFP concentration measurements (absorbance at 450 nm using an enzyme-linked immunosorbent assay) in various organs. sfGFP was detected at higher concentrations in the small intestine than in the stomach, both at extracellular and intracellular levels. Furthermore, sfGFP concentrations were higher in the lower small intestine compared to the upper small intestine, and the ratio of intracellular to extracellular concentrations was also increased. This result indicates that algal cells rupture within the small intestine, allowing sfGFP to be taken up by intestinal cells.

[0267] [Table 3]

[0268]

[0269] [Example 5]

[0270] (Dosage test and serum collection)

[0271] As the "control suspension administration group", 1.3 × 10 8 The sfGFP-expressing strain was suspended in 300 mM glucose solution (isotonic solution) at a concentration of 10 cells / mL (OD750 = 4). Using a probe, 300 μL of the suspension was delivered directly into the stomachs of three ICR mice. The same dose was administered orally six times every other week, and serum was collected two weeks after the final administration.

[0272] As the "suspension administration group", the 8 The Chl-TP-3HA-GP-Col expression strain (C. merolae strain expressing the ChlTP-sfGFP-HA-GP-Col protein) was suspended in 300 mM glucose solution at a concentration of 10 cells / mL (OD750 = 4). Using a probe, 300 μL of the suspension was delivered directly into the stomachs of four ICR mice. The same dose was orally administered six times every other week, and serum was collected two weeks after the final administration.

[0273] As the "alginate-solidified feed administration group", 27 mL of 300 mM glucose solution (OD750 = 4) in which the Chl-TP-3HA-GP-Col expression strain (ChlTP-sfGFP-HA-GP-Col protein expression strain of C. merolae) was suspended was centrifuged at 3000 g for 10 minutes, and the precipitated cells were collected. The cells of the Chl-TP-3HA-GP-Col expression strain and 1.12 g of commercial feed (CE-2) were suspended in 10 mL of a 2.5% sucrose solution containing 1% sodium alginate. Then, the suspension was dropped into a 10% calcium chloride solution to obtain a feed sample containing the Chl-TP-3HA-GP-Col expression strain and an alginate-solidified body of the commercial feed. The content of the Chl-TP-3HA-GP-Col strain was 4.6 mg wet weight / g (80 to 110 mg per pellet). Mice (four ICR individuals) were allowed to freely ingest the feed sample and then fed normally. The rats were fed the feed samples every other week for 6 times, and serum was collected 2 weeks after the final feeding.

[0274] (Evaluation of anti-GP protein antibody production)

[0275] The production of anti-GP protein antibodies was confirmed by immunoblotting. First, in order to fuse the 6× histidine tag sequence to the amino terminus of the GP protein of the rabies virus, the ORF of the GP gene was cloned into the pQE80 vector (containing a 6× histidine tag sequence, product number 32923, QIAGEN) to prepare a plasmid. The plasmid was introduced into Escherichia coli to express the 6× histidine tag fused GP protein (protein size is about 50kDa). The protein was concentrated using a nickel column (product number 17531901, GE Healthcare). Next, the 6× histidine tag fused GP protein concentrate was separated by electrophoresis using the SDS-PAGE method. The protein was transferred from the gel after electrophoresis to a polyvinylidene fluoride (PVDF) membrane (product number IPVH00010, Merck). The transferred membrane was immersed in a dilution of serum collected from each mouse individual and incubated at room temperature for 1 hour. Serum dilutions were prepared by diluting serum to 1 / 500 in Tris buffer (pH 7.5, containing 0.1% Tween 20). The presence of anti-GP protein antibodies in the serum was determined as the presence of antibody reaction with GP protein located around 50 kDa.

[0276] (result)

[0277] Figure 6 The results of immunoblotting are shown in FIG. In the serum dilutions of mouse individuals 1, 2, and 3 of the "control administration group (liquid)" as a negative control, no band was detected at the position of about 50 kDa, which is the molecular weight of the rabies GP protein ( Figure 6 (C)). In contrast, in the "alginate solidified feed administration group" mice 2, 3, and 4 ( Figure 6 (A)) and the “suspension administration group” mice 3 and 4 ( Figure 6 In (B), a band was detected at a position of approximately 50 kDa. This result indicates that mice administered with a suspension of C. merolae expressing rabies GP protein or an alginate-solidified diet produced anti-GP protein antibodies.

[0278] (Inspection)

[0279] In mouse individual 2 of the "control suspension administration group," a band smaller than the predicted size of the 6×histidine-tag fused GP protein was detected. This is believed to be due to the fact that the antibody possessed by this mouse individual reacted nonspecifically with the protein derived from Escherichia coli contained in the 6×histidine-tag fused GP protein concentrate, regardless of the administration of the GP protein.

[0280] A series of examples have confirmed that antigenic proteins appropriately introduced using the acid-resistant cells used in the present invention can be delivered to areas beyond the upper small intestine. Furthermore, it has been confirmed that even when the acid-resistant cells, which have been introduced with the antigenic proteins of the present invention, are mixed into feed in a form suitable for conventional livestock and aquaculture, the antigenic proteins can still be delivered to the target area. Furthermore, it has been confirmed that the antigenic proteins delivered in this manner stimulate the intestinal immune system, producing antibodies in the blood. Sequence Listing <110> Japan Science and Technology Agency <120> Drug Delivery Composition <130> PC-29322 <150> JP2019-069029 <151> 2019-03-29 <160> 66 <170> PatentIn version 3.5 <210> 1 <211> 2140 <212> DNA <213> Rabies virus <400> 1 acattttgag cctcttggat gtgaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 60 aaaaaaaaaa aaaaaactat taacatccct caaaagactt aaggaaagat ggttcctcag 120 gttcttttgt ttgcacccct cctggttttt ccattgtgtt tcgggaagtt ccccatttac 180 acgataccag acaaacttgg tccctggagc cctattgact tacaccatct cagctgtcca 240 aataacctgg ttgtggagga cgaaggatgt accaacctgt ccgggttctc ttacatggaa 300 cttaaagtgg gatacatctc agccataaa gtgaacgggt tcactgcac aggtgttgtg 360 acagaggcag aacctacac siactttgtt ggttatgtca caccacatt caagaaag 420 cattccgcc ccaccccaga cgcatgtaga gccgcgtata actggagat ggccggtgac 480 cccagatatg aagagtctct acacaatccg tacccgact accattggct tcgaactgta 540 aaaaccacca aagagtctct cgttatcata tcccaagtg tgacagattt ggacccatat 600 ɣaaatccc ttcactcaag gtcttccct ggcggaatt gctcaggaat aacggtgtcc 660 tcgacctact gctcacta tcatgattac accatctga tgctgaga tctgagacta 720 gggacatctt gtgacatttt taccatagc agaggagaga gagcatccaa aggagacaag 780 acttgcggct ttgtggatga aagaggcctg tataagtctt taaagggagc atgcaactc 840 aagttatgtg gagttctcgg acttagactt atggatggaa catgggtcgc gatgcaaca 900 tcagatgaga ccaatggtg ccctccaggt cagttggtga atttgcacga ctttcgctca 960 gacgagattg agcatctcgt tgaggagag ttagtcaaga aagagagga gtgtctgat 1020 gcactagagt ccatcatgac caccaagtca gtgagtttca gacgtctcag tcacctgaga 1140. aaacttgtcc ctgggtttgg aaaagcatat accatattca acaaaacctt gatggaggct gatgctcact acaagtctgt ccagacctgg aatgagatca tcccctcaaa agggtgtttg agagttgggg agaggtgtca tccccatgtg aacggggtgt ttttcaatgg our father 1260 gggtctgacg gccatgttct aatcccagag atgcagtcat ccctcctcca gcaacatatg 1320 gagttgttgg aatcttcagt tatccccctg atgcacccct tggcagaccc ttctacagtt 1380 ttcaaagacg gtgatgaggt tgaggatttt gttgaagttc acctccccga tgtgcataaa caggtctcag gagttgacct gggtctcccg aaatggggga agtatgtatt gatgattgca ggggccttga ttgccctgat gttgataatt ttcctgatga catgttgcag aagagtcaat cgaccagaat ctacacaaag caatcttgga gggacaggga gaaatgtgtc agtcccttcc caaagcgga aagtcatatc ttcatgggag tcatataaga gtggaggcga gaccagactg tgaaggccgg tcatcctttt gacacctcaa gtccagagga taacctcctc tcggggttgg ggggaatctt gggatccagt agtcctcctt gaactccatc caacagggta gatttaagag 1800 tcatgagact ttcattaatc atatcagttg atcagacatg gtcgtgtaga ttctcataac 1860 acgggagatc ttctagcagt ttcagtgacc aacggtgctt tcattctcca ggaactgata 1920 ccaaaggttg tggacaagcc aaggggtgct tcggattact ctgtgcttgg gcacagaaag 1980 aggtcatagt ttgccccttg atagcggatt caacatgaat taactaagaa aggcgatctg 2040 cctcccatga aggacataag caatagttca caatcatctt gcatctcagt gaagtgtaca 2100 taactataaa gggctgggtc atctaagcat ttcagtcgag 2140 <210> 2 <211> 524 <212> PRT <213> Rabies virus <400> 2 Met Val Pro Gln Val Leu Leu Phe Ala Pro Leu Leu Val Phe Pro Leu 1 5 10 15 Cys Phe Gly Lys Phe Pro Ile Tyr Thr Ile Pro Asp Lys Leu Gly Pro 20 25 30 Trp Ser Pro Ile Asp Leu His His Leu Ser Cys Pro Asn Asn Leu Val 35 40 45 Val Glu Asp Glu Gly Cys Thr Asn Leu Ser Gly Phe Ser Tyr Met Glu 50 55 60 Leu Lys Val Gly Tyr Ile Ser Ala Ile Lys Val Asn Gly Phe Thr Cys 65 70 75 80 Thr Gly Val Val Thr Glu Ala Glu Thr Tyr Thr Asn Phe Val Gly Tyr 85 90 95 Val Thr Thr Thr Phe Lys Arg Lys His Phe Arg Pro Thr Pro Asp Ala 100 105 110 Cys Arg Ala Ala Tyr Asn Trp Lys Met Ala Gly Asp Pro Arg Tyr Glu 115 120 125 Glu Ser Leu His Asn Pro Tyr Pro Asp Tyr His Trp Leu Arg Thr Val 130 135 140 Lys Thr Thr Lys Glu Ser Leu Val Ile Ile Ser Pro Ser Val Thr Asp 145 150 155 160 Leu Asp Pro Tyr Asp Lys Ser Leu His Ser Arg Val Phe Pro Gly Gly 165 170 175 Asn Cys Ser Gly Ile Thr Val Ser Ser Thr Tyr Cys Ser Thr Asn His 180 185 190 Asp Tyr Thr Ile Trp Met Pro Glu Asn Leu Arg Leu Gly Thr Ser Cys 195 200 205 Asp Ile Phe Thr His Ser Arg Gly Lys Arg Ala Ser Lys Gly Asp Lys 210 215 220 Thr Cys Gly Phe Val Asp Glu Arg Gly Leu Tyr Lys Ser Leu Lys Gly 225 230 235 240 Ala Cys Lys Leu Lys Leu Cys Gly Val Leu Gly Leu Arg Leu Met Asp 245 250 255 Gly Thr Trp Val Ala Met Gln Thr Ser Asp Glu Thr Lys Trp Cys Pro 260 265 270 Pro Gly Gln Leu Val Asn Leu His Asp Phe Arg Ser Asp Glu Ile Glu 275 280 285 His Leu Val Glu Glu Glu Leu Val Lys Lys Arg Glu Glu Cys Leu Asp 290 295 300 Ala Leu Glu Ser Ile Met Thr Thr Lys Ser Val Ser Phe Arg Arg Leu 305 310 315 320 Ser His Leu Arg Lys Leu Val Pro Gly Phe Gly Lys Ala Tyr Thr Ile 325 330 335 Phe Asn Lys Thr Leu Met Glu Ala Asp Ala His Tyr Lys Ser Val Gln 340 345 350 Thr Trp Asn Glu Ile Ile Pro Ser Lys Gly Cys Leu Arg Val Gly Glu 355 360 365 Arg Cys His Pro His Val Asn Gly Val Phe Phe Asn Gly Ile Ile Leu 370 375 380 Gly Ser Asp Gly His Val Leu Ile Pro Glu Met Gln Ser Ser Leu Leu 385 390 395 400 Gln Gln His Met Glu Leu Leu Glu Ser Ser Val Ile Pro Leu Met His 405 410 415 Pro Leu Ala Asp Pro Ser Thr Val Phe Lys Asp Gly Asp Glu Val Glu 420 425 430 Asp Phe Val Glu Val His Leu Pro Asp Val His Lys Gln Val Ser Gly 435 440 445 Val Asp Leu Gly Leu Pro Lys Trp Gly Lys Tyr Val Leu Met Ile Ala 450 455 460 Gly Ala Leu Ile Ala Leu Met Leu Ile Ile Phe Leu Met Thr Cys Cys 465 470 475 480 Arg Arg Val Asn Arg Pro Glu Ser Thr Gln Ser Asn Leu Gly Gly Thr 485 490 495 Gly Arg Asn Val Ser Val Pro Ser Gln Ser Gly Lys Val Ile Ser Ser 500 505 510 Trp Glu Ser Tyr Lys Ser Gly Gly Glu Thr Arg Leu 515 520 <210> 3 <211> 3033 <212> DNA <213> Cyanidioschyzon merolae (a red alga) <400> 3 atgttccatg tgacgtaccc gttcacgcag agacaatgct ttctccgttc acgagaagcg 60 tgccttgcaa cgttgccagc tggtgctttt cgaaagcacc tgtggcgccc ttcgtgctgg 120 tcgttccgca cacgtcttcg taaagaggcg tcgctacgga aatccacagt tctcgctccg 180 cttactcgcc gtctgcagct gagtctcttc ggcctcccag agcggttcgt tcgcaagtcc 240 aagtcgccgg tctcggcaga gtccagtgtc gccactgagc tcacacgtga tcgggtcaaa 300 gatccgacgc tcgcgaagta ctgggataca cttctggaaa tcaatgcact ggaggcggaa 360 ctggaacaac tcaaaagcga tgaactcaga gctcgcttgg atgccctgcg gagaaacgac 420 tcggtgcgga gcggggaccc gccactggcc gaggtattcg ccatcgttcg agaggccgca 480 cgtcggacgc tcagcatgcg acccttcgat gtgcaggttc ttggtggcct tgcactcttt 540 cacggttgcg tagcggagat cgccaccggt gaggggaaaa cgctcatcgc aacgatgccg 600 gcatgtgcca gcgcgctagc ggctcgcggt accgtcctgg tcgtgacggt gaacgattac 660 ctctgccgtc gcgactttga aaacatgggt ccactgtatc gctccctggg tttctctgtc 720 gggtgtgtga ccagcgccac agagcgggcg gcacgtcaac gagcatacgc ttgcgatatc 780 acctatgtga cgaatgcgga gcttggattc gactatctac gcgaccatct ggtgctgagc 840 gctgctgatc aagtgcttgt gaggcccaag cccttctact tttgtctact ggatgaggcc 900 gactcaatca tgatagatga agcgcgtaca ccgctgatca tttcccaggc tgcagaggcg 960 cccacagaga aatacgctac tgccgctaaa ctggctgcaa acctgcagcg ggatcggcac 1020 tacacggtct atgaaaagga gcgcaatgtc actttgacag gcgccggtta cgaagcatgt 1080 gaggaggcac tgcaagtgcc aacgctcttc gccgcagcgg atccgtgggc gccctttgtg 1140 ctgaatgcac tcaaggcgaa ggagctctat caacgtgata tagattatgt cgttcggggt 1200 gatcaagtgc taatcgtgga tgagtttacc ggtcgagtac tgcaaggtag gcgctggtca 1260 gagggtctgc accaggccat cgaggccaag gaggggctcg ctgtccgcac tgaaccgcgg 1320 actgtagctt ccatctcgta tcagtccttc tttcgcctgt ttcctcgtct ggcaggcatg 1380 acgggcaccg ctgctaccga tgcagcagag atacgcgaaa cgtacggact cgaggtggtc 1440 gttgtgccca ccgcgctacc tgtcgttcgc cgagactacc ccgatgtggt gtttcgaacg 1500 agtcgcggca aacttcttgc tgtggtcgca gaaattcgac gcctgcacct tcgaaaagtg 1560 cccgtcctgg ttggaaccac cagtattgaa gctagtgagc gaatcagcgc ccttttgagc 1620 gaaggcgaac gcgttccgca cgaggttctg aatgcacgtc cggagaacgc tgaacgtgag 1680 agcgaaatca tcgcccaagc aggtcgtcta ggagcggtta cgatcgcaac aaacatggct 1740 ggacgaggaa ccgatatcgt gctgggtgga aacgtgtcca gtctagcacg cgctcttctc 1800 cagagggagc tgcttgccac gttcgctctg gggcctgaat gctcctctgg ggccggcgac 1860 cgcgcatcca ccgagcattt cctctcgtgt ctcgaagagg cggagcatca ccagctgcac 1920 tgctttggag aaaaaatcgc tgaggcactg cgttcccagc gttcgacgga tcccggtccg 1980 cggcttatca tcgaatccat ggagcagctg catcagctga tgctgcaagc tgccgagttt 2040 caagagccaa cgttgccctt ctctgccgag gcccaggagc tggtaagaga ggcacttcag 2100 tgggctgaaa agcagctgcg tccgcggctc gatgcagagc gcgcagctgt gctggacctc 2160 ggtggcctgc acatcctcgg aactgaacga cacgagtcac gccgcatcga taaccagttg 2220 cggggtcgtg caggtcgcca aggtgacccc ggatgttcgc gtttcttcct gtcgctgggag 2280 gaccccatct tcagggtgtt tggggtgat cggatggctc gcctcgccga agcgtttcgt 2340 ctggacgaaa caacccccat cgagagcgtc caggttgcgc gtacgctgga caatgtccag 2400 cgcagcatcg agcaatatta tgcggggatt cggaagcagt tattcgctta tgatgaggtg 2460 ctttcccaac aacggaaggt actgtatcga cagcgaaacc gctttttgga agccgatgaa 2520 gcgctcctat tcggttcgga tgcggcagca gcccgggcct ctggggtgct tgctggcctc 2580 gcgggcgact ggatacgaac gacgattcag gatatctcc aagcaaatcg tcgagatccg 2640 gcaaaatgcg ttcagaaatt gaaagcgttc ttcccagggg cgcttctgaa cgagagcatg 2700 tgccagtccg cagcagctat cgaccaagtt gccgccgctg ttggcgtacg gctctcgcaa 2760 caccggcgga tgttgcaaca gagcgcaccg cagcaggatg ttgctgtctt tcgctatttg 2820 gcactggttc agcatgatca gctctggagt gaacatttac gaaaattagc gcttttgcgg 2880 gacatgtcgt ctttgcagac gttgcggcag gtagatccgt tgcaacagta tcagcaagat 2940 agttttcagc tctttgaaca gatgatggca caaataaggc gcaacactgt atactcgttt 3000 ttcaagtatt cgccggggcc aacggtatcc gcg 3033 <210> 4 <211> 1011 <212> PRT <213> Cyanidioschyzon merolae (a red alga) <400> 4 Met Phe His Val Thr Tyr Pro Phe Thr Gln Arg Gln Cys Phe Leu Arg 1 5 10 15 Ser Arg Glu Ala Cys Leu Ala Thr Leu Pro Ala Gly Ala Phe Arg Lys 20 25 30 His Leu Trp Arg Pro Ser Cys Trp Ser Phe Arg Thr Arg Leu Arg Lys<000079​​​​​​​​​ Lys Ser Pro Val Ser Ala Glu Ser Ser Val Ala Thr Glu Leu Thr Arg 85 90 95 Asp Arg Val Lys Asp Pro Thr Leu Ala Lys Tyr Trp Asp Thr Leu Leu 100 105 110 Glu Ile Asn Ala Leu Glu Ala Glu Leu Glu Gln Leu Lys Ser Asp Glu 115 120 125 Leu Arg Ala Arg Leu Asp Ala Leu Arg Arg Asn Asp Ser Val Arg Ser 130 135 140 Gly Asp Pro Pro Leu Ala Glu Val Phe Ala Ile Val Arg Glu Ala Ala 145 150 155 160 Arg Arg Thr Leu Ser Met Arg Pro Phe Asp Val Gln Val Leu Gly Gly 165 170 175 Leu Ala Leu Phe His Gly Cys Val Ala Glu Ile Ala Thr Gly Glu Gly 180 185 190 Lys Thr Leu Ile Ala Thr Met Pro Ala Cys Ala Ser Ala Leu Ala Ala 195 200 205 Arg Gly Thr Val Leu Val Val Thr Val Asn Asp Tyr Leu Cys Arg Arg 210 215 220 Asp Phe Glu Asn Met Gly Pro Leu Tyr Arg Ser Leu Gly Phe Ser Val 225 230 235 240 Gly Cys Val Thr Ser Ala Thr Glu Arg Ala Ala Arg Gln Arg Ala Tyr 245 250 255 Ala Cys Asp Ile Thr Tyr Val Thr Asn Ala Glu Leu Gly Phe Asp Tyr 260 265 270 Leu Arg Asp His Leu Val Leu Ser Ala Ala Asp Gln Val Leu Val Arg 275 280 285 Pro Lys Pro Phe Tyr Phe Cys Leu Leu Asp Glu Ala Asp Ser Ile Met 290 295 300 Ile Asp Glu Ala Arg Thr Pro Leu Ile Ile Ser Gln Ala Ala Glu Ala 305 310 315 320 Pro Thr Glu Lys Tyr Ala Thr Ala Ala Lys Leu Ala Ala Asn Leu Gln 325 330 335 Arg Asp Arg His Tyr Thr Val Tyr Glu Lys Glu Arg Asn Val Thr Leu 340 345 350 Thr Gly Ala Gly Tyr Glu Ala Cys Glu Glu Ala Leu Gln Val Pro Thr 355 360 365 Leu Phe Ala Ala Ala Asp Pro Trp Ala Pro Phe Val Leu Asn Ala Leu 370 375 380 Lys Ala Lys Glu Leu Tyr Gln Arg Asp Ile Asp Tyr Val Val Arg Gly 385 390 395 400 Asp Gln Val Leu Ile Val Asp Glu Phe Thr Gly Arg Val Leu Gln Gly 405 410 415 Arg Arg Trp Ser Glu Gly Leu His Gln Ala Ile Glu Ala Lys Glu Gly 420 425 430 Leu Ala Val Arg Thr Glu Pro Arg Thr Val Ala Ser Ile Ser Tyr Gln 435 440 445 Ser Phe Phe Arg Leu Phe Pro Arg Leu Ala Gly Met Thr Gly Thr Ala 450 455 460 Ala Thr Asp Ala Ala Glu Ile Arg Glu Thr Tyr Gly Leu Glu Val Val 465 470 475 480 Val Val Pro Thr Ala Leu Pro Val Val Arg Arg Asp Tyr Pro Asp Val 485 490 495 Val Phe Arg Thr Ser Arg Gly Lys Leu Leu Ala Val Val Ala Glu Ile 500 505 510 Arg Arg Leu His Leu Arg Lys Val Pro Val Leu Val Gly Thr Thr Ser 515 520 525 Ile Glu Ala Ser Glu Arg Ile Ser Ala Leu Leu Ser Glu Gly Glu Arg 530 535 540 Val Pro His Glu Val Leu Asn Ala Arg Pro Glu Asn Ala Glu Arg Glu 545 550 555 560 Ser Glu Ile Ile Ala Gln Ala Gly Arg Leu Gly Ala Val Thr Ile Ala 565 570 575 Thr Asn Met Ala Gly Arg Gly Thr Asp Ile Val Leu Gly Gly Asn Val 580 585 590 Ser Ser Leu Ala Arg Ala Leu Leu Gln Arg Glu Leu Leu Ala Thr Phe 595 600 605 Ala Leu Gly Pro Glu Cys Ser Ser Gly Ala Gly Asp Arg Ala Ser Thr 610 615 620 Glu His Phe Leu Ser Cys Leu Glu Glu Ala Glu His His Gln Leu His 625 630 635 640 Cys Phe Gly Glu Lys Ile Ala Glu Ala Leu Arg Ser Gln Arg Ser Thr 645 650 655 Asp Pro Gly Pro Arg Leu Ile Ile Glu Ser Met Glu Gln Leu His Gln 660 665 670 Leu Met Leu Gln Ala Ala Glu Phe Gln Glu Pro Thr Leu Pro Phe Ser 675 680 685 Ala Glu Ala Gln Glu Leu Val Arg Glu Ala Leu Gln Trp Leu Glu Lys 690 695 700 Gln Leu Arg Pro Arg Leu Asp Ala Glu Arg Ala Ala Val Leu Asp Leu 705 710 715 720 Gly Gly Leu His Ile Leu Gly Thr Glu Arg His Glu Ser Arg Arg Ile 725 730 735 Asp Asn Gln Leu Arg Gly Arg Ala Gly Arg Gln Gly Asp Pro Gly Cys 740 745 750 Ser Arg Phe Phe Leu Ser Leu Glu Asp Pro Ile Phe Arg Val Phe Gly 755 760 765 Gly Asp Arg Met Ala Arg Leu Ala Glu Ala Phe Arg Leu Asp Glu Thr 770 775 780 Thr Pro Ile Glu Ser Val Gln Val Ala Arg Thr Leu Asp Asn Val Gln 785 790 795 800 Arg Ser Ile Glu Gln Tyr Tyr Ala Gly Ile Arg Lys Gln Leu Phe Ala 805 810 815 Tyr Asp Glu Val Leu Ser Gln Gln Arg Lys Val Leu Tyr Arg Gln Arg 820 825 830 Asn Arg Phe Leu Glu Ala Asp Glu Ala Leu Leu Phe Gly Ser Asp Ala 835 840 845 Ala Ala Ala Arg Ala Ser Gly Val Leu Ala Gly Leu Ala Gly Asp Trp 850 855 860 Ile Arg Thr Thr Ile Gln Asp Ile Leu Gln Ala Asn Arg Arg Asp Pro 865 870 875 880 Ala Lys Cys Val Gln Lys Leu Lys Ala Phe Phe Pro Gly Ala Leu Leu 885 890 895 Asn Glu Ser Met Cys Gln Ser Ala Ala Ala Ile Asp Gln Val Ala Ala 900 905 910 Ala Val Gly Val Arg Leu Ser Gln His Arg Arg Met Leu Gln Gln Ser 915 920 925 Ala Pro Gln Gln Asp Val Ala Val Phe Arg Tyr Leu Ala Leu Val Gln 930 935 940 His Asp Gln Leu Trp Ser Glu His Leu Arg Lys Leu Ala Leu Leu Arg 945 950 955 960 Asp Met Ser Ser Leu Gln Thr Leu Arg Gln Val Asp Pro Leu Gln Gln 965 970 975 Tyr Gln Gln Asp Ser Phe Gln Leu Phe Glu Gln Met Met Ala Gln Ile 980 985 990 Arg Arg Asn Thr Val Tyr Ser Phe Phe Lys Tyr Ser Pro Gly Pro Thr 995 1000 1005 Val Ser Ala 1010 <210> 5 <211> 390 <212> DNA <213> Cyanidioschyzon merolae (a red alga) <400> 5 atgttccatg tgacgtaccc gttcacgcag agacaatgct ttctccgttc acgagaagcg 60 tgccttgcaa cgttgccagc tggtgctttt cgaaagcacc tgtggcgccc ttcgtgctgg 120 tcgttccgca cacgtcttcg taaagaggcg tcgctacgga aatccacagt tctcgctccg 180 cttactcgcc gtctgcagct gagtctcttc ggcctcccag agcggttcgt tcgcaagtcc 240 aagtcgccgg tctcggcaga gtccagtgtc gccactgagc tcacacgtga tcgggtcaaa 300 gatccgacgc tcgcgaagta ctgggataca cttctggaaa tcaatgcact ggaggcggaa 360 ctggaacaac tcaaaagcga tgaactcaga 390 <210> 6 <211> 130 <212> PRT <213> Cyanidioschyzon merolae (a red alga) <400> 6 Met Phe His Val Thr Tyr Pro Phe Thr Gln Arg Gln Cys Phe Leu Arg 1 5 10 15 Ser Arg Glu Ala Cys Leu Ala Thr Leu Pro Ala Gly Ala Phe Arg Lys 20 25 30 His Leu Trp Arg Pro Ser Cys Trp Ser Phe Arg Thr Arg Leu Arg Lys 35 40 45 Glu Ala Ser Leu Arg Lys Ser Thr Val Leu Ala Pro Leu Thr Arg Arg 50 55 60 Leu Gln Leu Ser Leu Phe Gly Leu Pro Glu Arg Phe Val Arg Lys Ser 65 70 75 80 Lys Ser Pro Val Ser Ala Glu Ser Ser Val Ala Thr Glu Leu Thr Arg 85 90 95 Asp Arg Val Lys Asp Pro Thr Leu Ala Lys Tyr Trp Asp Thr Leu Leu 100 105 110 Glu Ile Asn Ala Leu Glu Ala Glu Leu Glu Gln Leu Lys Ser Asp Glu 115 120 125 Leu Arg​​​​​​​​​​​​​​​​​​ ctctcaggtg aggcggtgga ggcgagtgca agcaaacgca agccccatct gaatgttggc 240 ggtatgggtc acgttgacca tgggaaaact acgctcgcgg cagcgattac gaaagtgctc 300 gccgagactg gtggagcccg gtacactgct tacgaagaga ttgacaaggc accggaggag 360 cgcgcccgcg ggatcacgat caacgcttcg catctcaaat acgagactcc atcacgttcg 420 tatgcacatg tcgattgccc tgggcatcga gactttgtga agaactttat cacgggtgcg 480 gcacaggtcg acaccgcgat tctcgtggtc agcggcccgg acggcccgca gccgcagact 540 caagagcacg tactgctatc gaagcaggta ggtgttccga actttgttgt atacctgaac 600 660 ctactcagca agtacgaata cgatggcgac aacgtgccga tcgtgcgggg ctctgccctg 720 aaggcattgc agggcgatca gagcgagctt ggctgtgggt ccatccacaa gctcctggag 780 attctcgaca aggtcccaat acccaaaaga gaccttgaaa aaccgttcct gatgcccatt 840 gaagatagct ttagcattac tggccgagga acggtggtta cgggacgcgt ggagaccggt 900 atcctacgcc ctggtgatga aatcgaaatc gtcggacttc gtcctcccga ggtagcacca 960 atgcgcacga tcgtcaccgg tatcgagact ttcaagcagt cgcttcctta cgcagaggct 1020 ggcgagaatg ttggctgtct gcttcgcggg gtcaagcgcg aagacgtgtt gcgcggtcag 1080 gttctagcga aaccgggaac ctccagagcg caccgcaagt tcgaggctga cgtttacatt 1140 cttactcagg aggaaggcgg ccgccataca ccattcttca gcaactatcg tcctcaattc 1200 ttcgtgcgga ctgcagatgt cacaggacgc ttccttctgc cgccagaagt ggagatgtgc 1260 atgccaggag atcgcgtacg atgcgctgtt gagctcatct atccggtcgc gctgcaggaa 1320 ggcttacgtt tcgctgttcg tgagggcggc aggaccgtcg gtgctgggtt ggttacgaaa 1380 gtcatcgag 1389 <210> 8 <211> 463 <212> PRT <213> Cyanidioschyzon merolae (a red alga) <400> 8 Met Ser Val Ser Cys Gly Arg Arg Ile Phe Ser Asp Ile Val Arg Gln 1 5 10 15 Val Arg Thr Phe Ala Thr Val Gly Val Asp Ala Arg Ser Leu Ala Gly 20 25 30 Thr Arg Asn Ala Thr Val Trp Arg Ser Tyr Arg Thr Thr Thr Leu Gln 35 40 45 Tyr Pro Arg Leu Trp Glu Leu Arg Ala Ser Arg Phe Leu Ser Gly Glu 50 55 60 Ala Val Glu Ala Ser Ala Ser Lys Arg Lys Pro His Leu Asn Val Gly 65 70 75 80 Gly Met Gly His Val Asp His Gly Lys Thr Thr Leu Ala Ala Ala Ile 85 90 95 Thr Lys Val Leu Ala Glu Thr Gly Gly Ala Arg Tyr Thr Ala Tyr Glu 100 105 110 Glu Ile Asp Lys Ala Pro Glu Glu Arg Ala Arg Gly Ile Thr Ile Asn 115 120 125 Ala Ser His Leu Lys Tyr Glu Thr Pro Ser Arg Ser Tyr Ala His Val 130 135 140 Asp Cys Pro Gly His Arg Asp Phe Val Lys Asn Phe Ile Thr Gly Ala 145 150 155 160 Ala Gln Val Asp Thr Ala Ile Leu Val Val Ser Gly Pro Asp Gly Pro 165 170 175 Gln Pro Gln Thr Gln Glu His Val Leu Leu Ser Lys Gln Val Gly Val 180 185 190 Pro Asn Phe Val Val Tyr Leu Asn Lys Cys Asp Met Val Asp Asp Pro 195 200 205 Glu Leu Leu Asp Leu Val Glu Leu Glu Val Arg Glu Leu Leu Ser Lys 210 215 220 Tyr Glu Tyr Asp Gly Asp Asn Val Pro Ile Val Arg Gly Ser Ala Leu 225 230 235 240 Lys Ala Leu Gln Gly Asp Gln Ser Glu Leu Gly Cys Gly Ser Ile His 245 250 255 Lys Leu Leu Glu Ile Leu Asp Lys Val Pro Ile Pro Lys Arg Asp Leu 260 265 270 Glu Lys Pro Phe Leu Met Pro Ile Glu Asp Ser Phe Ser Ile Thr Gly 275 280 285 Arg Gly Thr Val Val Thr Gly Arg Val Glu Thr Gly Ile Leu Arg Pro 290 295 300 Gly Asp Glu Ile Glu Ile Val Gly Leu Arg Pro Pro Glu Val Ala Pro 305 310 315 320 Met Arg Thr Ile Val Thr Gly Ile Glu Thr Phe Lys Gln Ser Leu Pro 325 330 335 Tyr Ala Glu Ala Gly Glu Asn Val Gly Cys Leu Leu Arg Gly Val Lys 340 345 350 Arg Glu Asp Val Leu Arg Gly Gln Val Leu Ala Lys Pro Gly Thr Ser 355 360 365 Arg Ala His Arg Lys Phe Glu Ala Asp Val Tyr Ile Leu Thr Gln Glu 370 375 380 Glu Gly Gly Arg His Thr Pro Phe Phe Ser Asn Tyr Arg Pro Gln Phe 385 390 395 400 Phe Val Arg Thr Ala Asp Val Thr Gly Arg Phe Leu Leu Pro Pro Glu 405 410 415 Val Glu Met Cys Met Pro Gly Asp Arg Val Arg Cys Ala Val Glu Leu 420 425 430 Ile Tyr Pro Val Ala Leu Gln Glu Gly Leu Arg Phe Ala Val Arg Glu 435 440 445 Gly Gly Arg Thr Val Gly Ala Gly Leu Val Thr Lys Val Ile Glu 450 455 460<00者: <210> 9 <211> 234 <212> DNA <213> Cyanidioschyzon merolae (a red alga) <400> 9 atgagcgtat cttgtggacg gcggatcttt tcagacattg tgcgtcaagt acgcacgttt 60 gctaccgtgg gcgtagacgc tcgctcgctt gcaggtacac gcaacgccac ggtctggcga 120 tcgtaccgca cgacgacgtt gcagtacccg cggctttggg aactgcgcgc ttcgcgcttc 180 ctctcaggtg aggcggtgga ggcgagtgca agcaaacgca agccccatct gaat 234 <210> 10 <211> 78 <212> PRT <213> Cyanidioschyzon merolae (a red alga) <400> 10 Met Ser Val Ser Cys Gly Arg Arg Ile Phe Ser Asp Ile Val Arg Gln 1 5 10 15 Val Arg Thr Phe Ala Thr Val Gly Val Asp Ala Arg Ser Leu Ala Gly 20 25 30 Thr Arg Asn Ala Thr Val Trp Arg Ser Tyr Arg Thr Thr Thr Leu Gln 35 40 45 Tyr Pro Arg Leu Trp Glu Leu Arg Ala Ser Arg Phe Leu Ser Gly Glu 50 55 60 Ala Val Glu Ala Ser Ala Ser Lys Arg Lys Pro His Leu Asn 65 70 75 <210> 11 <211> 690 <212> DNA <213> Cyanidioschyzon merolae (a red alga) <400> 11 atgggcgttc cgcaaatcac aagcaacacc gtggcacccg tcgaacgcaa ggtggagcgc 60 ggaagctacg gtgcaactgc gccagcagcg ccggcagctg cgttcgcctc caccgagtcc 120 gagctgatca gggatagtga cccttcgctc tcaacgatgt ggttgccggt ggatctgagt 180 gtccaactca ctccgaagga gcgtctgcag ctggcgtggt cggcggcacg gccgtggcga 240 gagtgggctg cactgcacgc actagcgccg ccgccaccct cgtcgtggct ggactggagc 300 gctcgtgtgc gcacgaattt ggagctctac gcatggaact acctctttgt ggcccttgtg 360 atgttcattg tgacgggttt gttctatccg tggagtgcgc tgctcctaat atcctggctc 420 ctgctcgcgc tgtacatggg tgtacgcacg gctgatgccg tcgggggcga ggacgcttcg 480 ctgggcgcgc gcatgttcca gaactggcct ggctatatcc gatacatttt gctgggtggc 540 ttgctcacgc tcatgttgtt tctgacagac gttgtcgcct tggcgctcac cagcgcatcg 600 ttggccgcag ctgtgacgct cgcgcacgca gcgctccacg atcccgttgc cgtgctctcc 660 gcgcggaatg ccgaatatac cgtggcaccg 690 <210> 12 <211> 230 <212> PRT <213> Cyanidioschyzon merolae <400> 12 Met Gly Val Pro Gln Ile Thr Ser Asn Thr Val Ala Pro Val Glu Arg 1 5 10 15 Lys Val Glu Arg Gly Ser Tyr Gly Ala Thr Ala Pro Ala Ala Pro Ala 20 25 30 Ala Ala Phe Ala Ser Thr Glu Ser Glu Leu Ile Arg Asp Ser Asp Pro 35 40 45 Ser Leu Ser Thr Met Trp Leu Pro Val Asp Leu Ser Val Gln Leu Thr 50 55 60 Pro Lys Glu Arg Leu Gln Leu Ala Trp Ser Ala Ala Arg Pro Trp Arg 65 70 75 80 Glu Trp Ala Ala Leu His Ala Leu Ala Pro Pro Pro Pro Ser Ser Trp 85 90 95 Leu Asp Trp Ser Ala Arg Val Arg Thr Asn Leu Glu Leu Tyr Ala Trp 100 105 110 Asn Tyr Leu Phe Val Ala Leu Val Met Phe Ile Val Thr Gly Leu Phe 115 120 125 Tyr Pro Trp Ser Ala Leu Leu Leu Ile Ser Trp Leu Leu Leu Ala Leu 130 135 140 Tyr Met Gly Val Arg Thr Ala Asp Ala Val Gly Gly Glu Asp Ala Ser 145 150 155 160 Leu Gly Ala Arg Met Phe Gln Asn Trp Pro Gly Tyr Ile Arg Tyr Ile 165 170 175 Leu Leu Gly Gly Leu Leu Thr Leu Met Leu Phe Leu Thr Asp Val Val 180 185 190 Ala Leu Ala Leu Thr Ser Ala Ser Leu Ala Ala Ala Val Thr Leu Ala 195 200 205 His Ala Ala Leu His Asp Pro Val Ala Val Leu Ser Ala Arg Asn Ala 210 215 220 Glu Tyr Thr Val Ala Pro 225 23o <210> 13 <211> 1287 <212> DNA <213> Cyanidioschyzon merolae (a red alga) <400> 13 atggtgatca gcacccaggg aaacgtctca ggttccgttc cgaatacgct cagagcgaaa 6o cctgagcaaa cgcttcaggc gtcgaggcct gccaccaggg ccgtctctgc agcgcttgac 120 ttctgcgctg gagttgcagc agccgtttca ggtatatctg ttcccggggg atattttccc 180 It should be noted that there is a possible error in the original text where "225 23o" should probably be "225 230". This has been corrected in the translation as much as possible while maintaining the integrity of the original text.cctttcatgt cattcagcaa ccgcttaagt ccaaaagctg gcacagatac tctttcggtg agcaatcagg agtcaaccag ggaagacgcg gacgctgtgc tttcgacggc agccgcttcg ctggcaacgg acctcgagta cggctccgcg acagagaggc accacgaga catcgatcct 360 attcgcccag tgacgcgaga ctctggcgca ttcgtcgaga actatccag tggaacacgg 420 gaggtggcaa tcgctttcga aggagtgacg ttgtctgcga gcacccaggc ggcggggacg 480 actcgaccca tacttcggaa tatctgtttc gaagtacgcg acggagagac ggtgtttatc atcggcccgt caggagctgg caagtctcga cttcttcgac tggtgaaccg cttagaggag ccttcaggtg gtcaagtccg gctgtggggc acaccagtgc cagcgtaccc tccaggcgag 660 ttacggggca aactagtag ttttctgagc cagcaacctg cattgccgtg tttggcacat 720 aggaggcctg gtttgctgga ggcactccgc cgacttgtaa cgcgagaaac gctttgggag 780 ctggtcctcg gcaaacgccg acaacgctcg agcgcgaca cggaggacc agcaaagtcg 840 gcgctcgaga cgctcgttcg tttggggagtt gtatcgcgga cagagcttga acagcgacta ccggaagcac tgcatatagc tggcttatcg cgaacgattc tggatcgacc gctagccgcc 960 ttgtcagggg gtgagcgagc tcgcctcggc ctcacgcgag tgcttttgca gcaaccacga 1020 atcctactgc tggacgaggt cacttcaagt ctggacgcag caacagcggc tcaagtattg 1080 caacgtttat cggactggaa agaacgcatc cgggccacaa tactcatagt gacgcacaga 1140 ctctcggagg ttaccgacgg ccagctcatt cttgtgcggg acggggagct gttcttacgc 1200 ggtgacgcgc gtgcgctgct cggtaccgcc gatacagcgg ccagcattca tcggcttttg 1260 actggggagt cattgggagc aacaaaa 1287 <210> 14 <211> 429 <212> PRT <213> Cyanidioschyzon merolae (a red alga) <400> 14 Met Val Ile Ser Thr Gln Gly Asn Val Ser Gly Ser Val Pro Asn Thr 1 5 10 15 Leu Arg Ala Lys Pro Glu Gln Thr Leu Gln Ala Ser Arg Pro Ala Thr 20 25 30 Arg Ala Val Ser Ala Ala Leu Asp Phe Cys Ala Gly Val Ala Ala Ala 35 40 45 Val Ser Gly Ile Ser Val Pro Gly Gly Tyr Phe Pro Pro Phe Met Ser 50 55 60 Phe Ser Asn Arg Leu Ser Pro Lys Ala Gly Thr Asp Thr Leu Ser Val 65 70 75 80 Ser Asn Gln Glu Ser Thr Arg Glu Asp Ala Asp Ala Val Leu Ser Thr 85 90 95 Ala Ala Ala Ser Leu Ala Thr Asp Leu Glu Tyr Gly Ser Ala Thr Glu 100 105 110 Arg His His Glu Asn Ile Asp Pro Ile Arg Pro Val Thr Arg Asp Ser 115 120 125 Gly Ala Phe Val Glu Asn Tyr Pro Ser Gly Thr Arg Glu Val Ala Ile 130 135 140 Ala Phe Glu Gly Val Thr Leu Ser Ala Ser Thr Gln Ala Ala Gly Thr 145 150 155 160 Thr Arg Pro Ile Leu Arg Asn Ile Cys Phe Glu Val Arg Asp Gly Glu 165 170 175 Thr Val Phe Ile Ile Gly Pro Ser Gly Ala Gly Lys Ser Arg Leu Leu 180 185 190 Arg Leu Val Asn Arg Leu Glu Glu Pro Ser Gly Gly Gln Val Arg Leu 195 200 205 Trp Gly Thr Pro Val Pro Ala Tyr Pro Pro Gly Glu Leu Arg Gly Lys 210 215 220 Leu Val Gly Phe Leu Ser Gln Gln Pro Ala Leu Pro Cys Leu Ala His 225 230 235 240 Arg Arg Pro Gly Leu Leu Glu Ala Leu Arg Arg Leu Val Thr Arg Glu 245 250 255 Thr Leu Trp Glu Leu Val Leu Gly Lys Arg Arg Gln Arg Ser Ser Ala 260 265 270 Asn Thr Glu Glu Pro Ala Lys Ser Ala Leu Glu Thr Leu Val Arg Leu 275 280 285 Gly Val Val Ser Arg Thr Glu Leu Glu Gln Arg Leu Pro Glu Ala Leu 290 295 300 His Ile Ala Gly Leu Ser Arg Thr Ile Leu Asp Arg Pro Leu Ala Ala 305 310 315 320 Leu Ser Gly Gly Glu Arg Ala Arg Leu Gly Leu Thr Arg Val Leu Leu 325 330 335 Gln Gln Pro Arg Ile Leu Leu Leu Asp Glu Val Thr Ser Ser Leu Asp 340 345 350 Ala Ala Thr Ala Ala Gln Val Leu Gln Arg Leu Ser Asp Trp Lys Glu 355 360 365 Arg Ile Arg Ala Thr Ile Leu Ile Val Thr His Arg Leu Ser Glu Val 370 375 380 Thr Asp Gly Gln Leu Ile Leu Val Arg Asp Gly Glu Leu Phe Leu Arg 385 390 395 400 Gly Asp Ala Arg Ala Leu Leu Gly Thr Ala Asp Thr Ala Ala Ser Ile 405 410 415 His Arg Leu Leu Thr Gly Glu Ser Leu Gly Ala Thr Lys 420 425 <210> 15 <211> 1362 <212> DNA <213> Cyanidioschyzon merolae (a red alga) <400> 15 atgtcgtcca tctgggagca aatgagatct ctgctgttcg gagcgagtgg atcgtatgct 60 gatgcggccg acacacccca agggcttgct ttcgaacagc gacttgttgc gaggcacgga 120 aacacgctgc cgccttggcc gctcttcaag ctgggtgctg tatacggcgt tggaccagtg 180 cctcccaacc tgcgggccct tctagacggt tctttctttc tgataggcgc gtgtttgggg 240 actgcggcgc tgtttgactt tgccggtatc atcggtagac gcggcggcgg cgttactgca 300 gcggaagttg cgaaagagtg caactgtgcc gatgtcaatg cggtcggtcg cgttctacgg 360 gcttgcgaaa actggggcta ctttgagtcc tacgtcccgc gggactgtga ctcaaagagc 420 gtatcgaacg agcagcgctt atggcggaac acactattat cggctttatt gcgcgaggat 480 catccacaca gcgtgcgcgc tcaaataatg cacttgtacg tcgatatttt tcccgcgagt 540 gcgcttctct ttgagacgat acgtgatacg ccgtcaaacg acgtagaaac ggacggtttg 600 cgagcgcgtg ccgatgcgtc agcgacaaaa caaccgagcg cgtttgaacg cgtccatcag 660 tgtacgtttt gggaatacct gtcgagccat ccggatcgat gggatgtctt caacagggcc 720 atgcgtagct cagatgcgct cctgggatcg acgatcatga aagatatcga ctggggacgc 780 tacctccggg ttatagacct cggcgcagcg gatggttcac tcgtttatca tctgctgagc 840 gctttcgcgc tgaaggcagt gattttcgat ctgccgccgg ttatcgagca tgccaaggcg 900 tactggaaca cgagtccgga gcgctcggca atggtcgcaa gtggccgcgt ccagttcgcg 960 gccggcgacc ttttcgacgc caccaccgtg ccagcagcgg aggaaggcga catctatgtg 1020 atgcgcaata tctggcacga ctggcgcgat cccgactgta tccgcattgg ccggagtgtt 1080 cgagcagcaa tcggcgacgt tcgcaatgtg aagctcgtca tcctggaggc aagcattgac 1140 cagtatccgc gcggctcact ccttgagcgc tttcgcgttg cactcgatca aatcatgttt 1200 accgcatttc gatccaagga acgcgacagg atagagtttg atgcgctctt acgtcgatgt 1260 ggttttcagc tgaccgaagt gaggcattta cgcgcatcac tcgtcgcagt gatcgccgag 1320 ccgctttcgc attgggtcca gtctcctgaa ccagcggacc aa 1362 <210> 16 <211> 454 <212> PRT <213> Cyanidioschyzon merolae (a red alga) <400> 16<000124l>Met Ser Ser Ile Trp Glu Gln Met Arg Ser Leu Leu Phe Gly Ala Ser 1 5 10 15 [[ID=z6]]Gly Ser Tyr Ala Asp Ala Ala Asp Thr Pro Gln Gly Leu Ala Phe Glu 20 25 30 Gln Arg Leu Val Ala Arg His Gly Asn Thr Leu Pro Pro Trp Pro Leu 35 40 45 Phe Lys Leu Gly Ala Val Tyr Gly Val Gly Pro Val Pro Pro Asn Leu 50 55 60 Arg Ala Leu Leu Asp Gly Ser Phe Phe Leu Ile Gly Ala Cys Leu Gly 65 70 75 80 Thr Ala Ala Leu Phe Asp Phe Ala Gly Ile Ile Gly Arg Arg Gly Gly 85 90 95 Gly Val Thr Ala Ala Glu Val Ala Lys Glu Cys Asn Cys Ala Asp Val 100 105 110 Asn Ala Val Gly Arg Val Leu Arg Ala Cys Glu Asn Trp Gly Tyr Phe 115 120 125 Glu Ser Tyr Val Pro Arg Asp Cys Asp Ser Lys Ser Val Ser Asn Glu 130 135 140 Gln Arg Leu Trp Arg Asn Thr Leu Leu Ser Ala Leu Leu Arg Glu Asp 145 150 155 160 His Pro His Ser Val Arg Ala Gln Ile Met His Leu Tyr Val Asp Ile 165 170 175 Phe Pro Ala Ser Ala Leu Leu Phe Glu Thr Ile Arg Asp Thr Pro Ser 180 185 190 Asn Asp Val Glu Thr Asp Gly Leu Arg Ala Arg Ala Asp Ala Ser Ala 195 200 205 Thr Lys Gln Pro Ser Ala Phe Glu Arg Val His Gln Cys Thr Phe Trp 210 215 220 Glu Tyr Leu Ser Ser His Pro Asp Arg Trp Asp Val Phe Asn Arg Ala 225 230 235 240 Met Arg Ser Ser Asp Ala Leu Leu Gly Ser Thr Ile Met Lys Asp Ile 245 250 255 Asp Trp Gly Arg Tyr Leu Arg Val Ile Asp Leu Gly Ala Ala Asp Gly 260 265 270 Ser Leu Val Tyr His Leu Leu Ser Ala Phe Ala Leu Lys Ala Val Ile 275 280 285 Phe Asp Leu Pro Pro Val Ile Glu His Ala Lys Ala Tyr Trp Asn Thr 290 295 300 Ser Pro Glu Arg Ser Ala Met Val Ala Ser Gly Arg Val Gln Phe Ala 305 310 315 320 Ala Gly Asp Leu Phe Asp Ala Thr Thr Val Pro Ala Ala Glu Glu Gly 325 330 335 Asp Ile Tyr Val Met Arg Asn Ile Trp His Asp Trp Arg Asp Pro Asp 340 345 350 Cys Ile Arg Ile Gly Arg Ser Val Arg Ala Ala Ile Gly Asp Val Arg 355 360 365 Asn Val Lys Leu Val Ile Leu Glu Ala Ser Ile Asp Gln Tyr Pro Arg 370 375 380 Gly Ser Leu Leu Glu Arg Phe Arg Val Ala Leu Asp Gln Ile Met Phe 385 390 395 400 Thr Ala Phe Arg Ser Lys Glu Arg Asp Arg Ile Glu Phe Asp Ala Leu 405 410 415 Leu Arg Arg Cys Gly Phe Gln Leu Thr Glu Val Arg His Leu Arg Ala 420 425 430 Ser Leu Val Ala Val Ile Ala Glu Pro Leu Ser His Trp Val Gln Ser 435 440 445 Pro Glu Pro Ala Asp Gln 450 <210> 17 <211> 1488 <212> DNA <213> Cyanidioschyzon merolae (a red alga) <400> 17 atggacccaa ccaagtatac accacgcgat acttttgacg tgaacgtgct taccacgaac 60 gctgggcagc cggtgacgaa caaccagtct tcacgcactg tggggccgcg tggaccggtg 120 ctgcttgagg actaccacct cttggagaag ctggctaact ttgaccgtga acggcaaccg 180 gagcgtgttg tacacgcgcg cggtgtgact gccaagggtt actttgaggt gacacacgat 240 atcacagact taacctgcgc ggacatgttc gcggaggttg gtcgccgtac gccggtggct 300 gtccggtttt cgacggtcat tcactcgcgt cattcaccgg aaaccttgcg tgatccgcgc 360 ggttttgctg tcaagttcta cactcgtgaa ggaatctggg acctcgtcgg aaataatctg 420 ccggtcttct ttatcaggga tgcgatcaag ttcccggact taattcacgc gttcaaacca 480 aacccgcgga cagaggcgca ggaatcctgg aggattctcg actttttaag caaccaacac 540 gaaagcctga atatgctcac gttccttttc gacgacgagg gtatcccgaa ggactatcgg 600 catatgcgcg gcagcggagt gcactcgttc cgccttgtaa ccagggacgg acgctcgacg 660 tacgtgcgct tccactggcg ccccaagtgc ggtatggaaa acttgttgga cgaagaggct 720 gccgttgtgt gtggtcagga tttttctcac gcaacgcatg acttgatccg ggcaattgac 780 cgaggcgact atcccgaatg ggcgctctat atccaaacca tggacccggc aatggttgag 840 aaccacgtgt tcccgtgggg cgatccactc gacgcgacga gagagtggcc tgagaaggac 900 tttccgcttc gccctgtggg tcgcatggtg ctgaatcaga actgcgataa ccagttcttg 960 gagaatgagc aaattgcctt ctcgccggca ctcgttgtcc caggtatcta ctattcagac 1020 gataagctgt tgcagggtcg ccttttcagt tacgccgaca cgcagcgata ccgtattggt 1080 gccaattatc tgcagctgcc gatcaacgca ccaaagaatc cgttccataa taaccattat 1140 gatggccagc agaactggat gctgcgtcag ggcgaagtga actactaccc cagccgcgta 1200 gacccggtgc cggaggcgcc cccggccgca ttcccaacgc ctcgcgatga actacgtggt 1260 cagcgggtga agcagctggt tccaaatcag tgcgactttg tgcagcctgg agaacgttac 1320 cgctctttcg atccggcgcg caaggagcgg ttcgtgaacc ggatcgcgaa actgctaaca 1380 gatgagcgcg tgaccccaga gctgcgagcc atctggctgg agctctggag caaatgcgat 1440 gctgaactgg gcgcagcgct ggcgaccaga gtgaagcagt gtacaatg 1488 <210> 18 <211> 496 <212> PRT <213> Cyanidioschyzon merolae (a red alga) <400> 18 Met Asp Pro Thr Lys Tyr Thr Pro Arg Asp Thr Phe Asp Val Asn Val 1 5 10 15 Leu Thr Thr Asn Ala Gly Gln Pro Val Thr Asn Asn Gln Ser Ser Arg 20 25 30 Thr Val Gly Pro Arg Gly Pro Val Leu Leu Glu Asp Tyr His Leu Leu 35 40 45 Glu Lys Leu Ala Asn Phe Asp Arg Glu Arg Gln Pro Glu Arg Val Val 50 55 60 His Ala Arg Gly Val Thr Ala Lys Gly Tyr Phe Glu Val Thr His Asp 65 70 75 80 Ile Thr Asp Leu Thr Cys Ala Asp Met Phe Ala Glu Val Gly Arg Arg 85 90 95 Thr Pro Val Ala Val Arg Phe Ser Thr Val Ile His Ser Arg His Ser 100 105 110 Pro Glu Thr Leu Arg Asp Pro Arg Gly Phe Ala Val Lys Phe Tyr Thr 115 120 125 Arg Glu Gly Ile Trp Asp Leu Val Gly Asn Asn Leu Pro Val Phe Phe 130 135 140 Ile Arg Asp Ala Ile Lys Phe Pro Asp Leu Ile His Ala Phe Lys Pro 145 150 155 160 Asn Pro Arg Thr Glu Ala Gln Glu Ser Trp Arg Ile Leu Asp Phe Leu 165 170 175 Ser Asn Gln His Glu Ser Leu Asn Met Leu Thr Phe Leu Phe Asp Asp 180 185 190 Glu Gly Ile Pro Lys Asp Tyr Arg His Met Arg Gly Ser Gly Val His 195 200 205 Ser Phe Arg Leu Val Thr Arg Asp Gly Arg Ser Thr Tyr Val Arg Phe 210 215 220 His Trp Arg Pro Lys Cys Gly Met Glu Asn Leu Leu Asp Glu Glu Ala 225 230 235 240 Ala Val Val Cys Gly Gln Asp Phe Ser His Ala Thr His Asp Leu Ile 245 250 255 Arg Ala Ile Asp Arg Gly Asp Tyr Pro Glu Trp Ala Leu Tyr Ile Gln 260 265 270 Thr Met Asp Pro Ala Met Val Glu Asn His Val Phe Pro Trp Gly Asp 275 280 285 Pro Leu Asp Ala Thr Arg Glu Trp Pro Glu Lys Asp Phe Pro Leu Arg 290 295 300 Pro Val Gly Arg Met Val Leu Asn Gln Asn Cys Asp Asn Gln Phe Leu 305 310 315 320 Glu Asn Glu Gln Ile Ala Phe Ser Pro Ala Leu Val Val Pro Gly Ile 325 330 335 Tyr Tyr Ser Asp Asp Lys Leu Leu Gln Gly Arg Leu Phe Ser Tyr Ala 340 345 350 Asp Thr Gln Arg Tyr Arg Ile Gly Ala Asn Tyr Leu Gln Leu Pro Ile 355 360 365 Asn Ala Pro Lys Asn Pro Phe His Asn Asn His Tyr Asp Gly Gln Gln 370 375 380 Asn Trp Met Leu Arg Gln Gly Glu Val Asn Tyr Tyr Pro Ser Arg Val 385 390 395 400 Asp Pro Val Pro Glu Ala Pro Pro Ala Ala Phe Pro Thr Pro Arg Asp 405 410 415 Glu Leu Arg Gly Gln Arg Val Lys Gln Leu Val Pro Asn Gln Cys Asp 420 425 430 Phe Val Gln Pro Gly Glu Arg Tyr Arg Ser Phe Asp Pro Ala Arg Lys 435 440 445 Glu Arg Phe Val Asn Arg Ile Ala Lys Leu Leu Thr Asp Glu Arg Val 450 455 460 Thr Pro Glu Leu Arg Ala Ile Trp Leu Glu Leu Trp Ser Lys Cys Asp 465 470 475 480 Ala Glu Leu Gly Ala Ala Leu Ala Thr Arg Val Lys Gln Cys Thr Met 485 490 495 <210> 19 <211> 8157 <212> DNA <213> Cyanidioschyzon merolae <400> 19 atgggggatt tgggtgaaca tgcaccgtcg aatggctccg caaaggatag cgttcggcgc tatgttgagg aacacggcgg tagtagcca attcagcgct tattggtcgc aaacaacgga atcgccgccg taaagtgcat ccgatcaatg cgaagatggg catacgaagc gtttggtagt gaacgagcgc tggagtttgt agcaatggcc actcccgagg atgttcaagc caacgccgac 300. 300. cttcgcc tcgctgacct cttcgttccc gtacctgggg gccaacgtgg agctcatcgt cgacatcgcg gaaaggaacg agtgcgatgc agtatgggcg 360 ggttggggtc acgccagcga gaaccccgctg ctgcccgctc gccttgcgga gactggtatc 420 gcattcctgg gaccggatgc cattgccatg agggcgctcg gcgacaagat ttcgtcaacg 480 ttgttagctc agtctgcgga tgttccagtc gtcagctgga acggtgatga cttgaaggta 540 acttttcacc gcgagcgcgg cggtattgat gaagaacgct atcgccgtgc ctgcgttgcg 600 aatgtcgacg aggctcgcgc tgctgccgat cgtattggct acccagtgat gattaaagcc agcgaaggag gcggtggcaa aggcatccgt ctttgccgtc gaccagagga tgttcgagat 720 gcctttcgtc aggtcgccgg agagattcct ggctctccga tcttcatcat gaaaatggtg 780 gatcaagcgc gacacctgga agtgcagatt gttgccgatg aatacgggca cgcgatagcg 840 ctatatgggc gtgactgctc cgtgcagaga cggcatcaaa agatcatcga ggaagggcca 900 gttacggcaa cgcaccgca ggtttggaaa gagctcgaac atggagcggt gcgtcttgcc 960 aaaatggtcg gttacgtagg tgccggcact gttgagtatc tctacgacgg tcgacgtttc 1020 tatttcctgg aactgaatcc gcgcttgcag gtggagcacc cagtaaccga gtggatcact 1080 ggcgtgaacc tgcctgcagt ccagctgcag attgccatgg ggattccgct ccagcgtatt 1140 gcagctatcc aagcacttta cggcaataga ccggacctgg atctcgagcg acaccagccg 1200 aacccgccgc atggccacgt aatcgcctgc cgcgttaccg cagaaaatcc cgagaagga 1260 ttccagccga caagcggctc cattcaggag ctttcgtttc gaaatacgcc caatgtgtgg 1320 ggctatttca gcgtcggcgc ctggggtggt gtccacgagt acgctgactc gcaattcggg 1380 cacctttttg cctggggtgc ggatcgcgag atggcccgga ggaatatggt gctggcactg 1440 aaagagctca gcatccgcgg agatatccgc acaacggtgg agtatcttat tacgttgctg 1500 gagctcgaat cgtatcgcga aaatcgcatc catacccgtt ggctggacaa cctcattgcc 1560 tccaaagtga aaccagagcg accgccgttc catatcgctg ttgtgcttgg tgcagtccat 1620 caggcgtaca gagcctggtc tgagcgccgg cgatcttttg ttgagtcgct gagtcgcggt 1680 caagtgccgc agcgtattga cgctaccttc atagaatttc agttcgagct gatctacgat 1740 gagctcaagt acgcgctcat cgtcaggcaa gcgggtccga atgccttcca cgtctcccta 1800 gcacagcatc ccgaggaacg ggtgcgtgtt gatttccgac ctctgctgga cggtggtctc 1860 ttgatcatgt gcgatggtcg ctccttcaat actcactcca aggaggattc cactggcatg 1920 cgtctgagca tcgacgggcg aacctgtgtg ttcccaggcg aaatggaccc tacgcgcatt 1980 gcagcacagg cctcggggcg tctcgtccga tacctggttg ccaacgagga acacgtcgat 2040 cagggacaag tcattgcgga gattgaagtc atgaaaatgt atctgagcgt gcaggctccc 2100 gaaccgggca ccattcactt gctgaaacca gcgggagcgg ccctggagcc aggcgaagtt 2160 tttgcgcagc tcgatctcga cgatccgagc aaagttcgac gggtgacgcc atttactggg 2220 cgctttccga caatgctgcc tccacagcga ctgggaaaga aaccgcacca gcgctttgag 2280 agcgcgaaac agcagatcga agcgctcctg gacggctacg atgttgatat ggagccgctc 2340 ggcatgcttc tgcagctggc gagcaccgaa ccggcagtac cggccgggaa actgcaggag 2400 gcgctctctt tcctagccgg tcgtattccg tcgacgttgc atgccgccct tctaggacaa 2460 gttttcgaac tgcgttccgt tgctcgtgat gatcacgagc gcctccaaca gcatttccgc 2520 gaaatgaagc gcatgatcag cgatttctgc gagcaacttg ccatcgcagc ggaccgcgag 2580 gcgctggagg ctaccctgcg ccccctgacc agtatcctgg aggcgtatgt tcgcgggggc 2640 cttcgcggct accacgagca tctcgttgta tgcctcatgc aacgctatgt gaatacggag 2700 aagtattttg cgcaaggtcg gcggctcgat gaggttctat tcgatctgcg cgaacaacac 2760 cgcgagcact tggaagttgt tgcggatatc atgctcgcac atgcccagtt agcgcgaaag 2820 aaccaactga tgctcgcttt gctcgatcac atagccgcgg atgcatcgct tctgcgcagt 2880 gtcattgtcc ggcagtgcct tcacgaggtg gctgcgttca tccaccccga ctatagccaa 2940 ctggcgctgc gagcaagact cttgctggcg agttctcgtc gccgagcgct ccccgaacgt 3000 cgagagcgcc tctgcaagca gatcgaacag gctgtacatg cgcccactga agaacaatgc 3060 tatcgtctgt tgcacgcggt ggtcgagtcc caggaaagca tcctggatgc gctggtgagc 3120 ctaaccatga acccgggggt gccggtggag attcgcaaag ccgctgtcca gtgccagatc 3180 ctgcgggcct acaaagcgta ccacgtgtac gatctggttg tcgaacttga cgaggaactg 3240 ggtttcctgc gggcgttgtg gcgtttccag tatcgttcaa acctgaacgc ttttggtagt 3300 cattccagta tgcggtcgtt tccggcggca ttggtggcgg cgtcggctcc gctagcccgc 3360 cggcagctgc gtagctacga ctctgcggac aatttgcaga actggggctt tcgagtcggt 3420 atgttggtcg tctttgagac gctgcgggcg atggtgcagg gtttcgatcg tgtgctccag 3480 gtttttcgag cggaaaccgg cggcgaaacg ctggcttcgc ctcgtaccaa gtcgagcgcc 3540 gcagtcgatt ccagtgcaga ctccagtgct gaaggtattg gtgtcaatgt actgaccatc 3600 gccattccct ggacggccgt ggaggtcgct gattttgcga aatacctggg cgttgcagac 3660 gcggacgacc gtctgcaggc aagtaatgcg aacgactcgg agcaggttgt cgccctgctc 3720 acgcgtttct gtcgctcttc cgcggagcgc aaagcctcga tgcgcgctgc cggtatcaaa 3780 cttgtgacct ttttggtggc acccggtgaa ctgtgccagc gcagctggg ctccagtctg 3840 cctcgtgaaa gcacttatcc gggtttctat acgttgcggg catcgctcga gtacgccgag 3900 gatcccattt atcgccacat tgatccacca gcggctttcc agctggagct gaatcgattg 3960 gcgaactttc gtatcacccg ctttgaacat ccgaaccgct ccattcacgt cttctacgcg 4020 gaagatcgaa ccgacaaggg cgacgctcgc ttcttcgtgc gcgcttttgt acggcaagcg 4080 gaagtctacg ccagtcccag cgatacagca gcggtttcga tcccggaagc ggagaggact 4140 tttgtggcct gtctggatgc cttggaaacc gcccgttgtg ataggcgctt ccgacgaacg 4200 gactttaatc atctcttctt gcacctgatt ccccgggtct ccatcgatgt ggatgacgtc 4260 gaggcgatct gtacgcgcct cttttatcgg ttctcctcac gttgctggcg tttgcgcgtg 4320 ttcatggtag agatcaaagt ccatgtggaa aaaatgggcc cgaaaccgct tcgatttatt 4380 ctctataatc caaccggaca ctcgttacgc gttgaaggct acgtggaaca gggagatcgt 4440 ctcctgtcgt tggattccag cgatccgggt catttacatg gtacacccgt cgatgagccg 4500 taccaggtac tgaaccgtat ccagcgccgg cgagttgttg cccagacgct cgaaacaact 4560 tacgtctacg attaccagga gcttttcacc aaagcactcc atgagcagtg gcggcgttat 4620 tcgcaggaac gacttttgg aggatttcgc cgccacaaga taccgctgaa actgctgacc 4680 tgcacggagc tcgttctcca ggatgaagat catgacgaga gcgagctgat agagaccaac 4740 cgccccgccg gggagaatga aatcggtatg gtcgcctggc gctacacctt cttcacgcccc 4800 gagtatcccc agggtcgcga tgcgatcgtc attgccaacg atatcacgta cctgagtgga 4860 tcgtttggac ctcgagagga tcgcctcttt gccaaggcgt ctcagcaggc gagaaagctc 4920 ggtataccgc gcatctacat cgctgcgaat agcggagccc gaatcggcct cgcggaagaa 4980 ctgcgcaacg tcttccaagt tcagtggaag gatgcacatg acccgagccg aggcttcgac 5040 tacctctacc tgggactggc ggataagctc aagtacgagg atgaactcgg catcgtgcga 5100 acgcgagacg tcggcggcgg caagtacgct ctagttgata tctatggcgc tgaggatggc 5160 attggtgtcg agaacctcat gggttcagcg tgcatcgctg ccgagacctc agccgcctac 5220 aatgactgct tcaccatcac gtacgtcgca gcgcgctgtg tggggatcgg tgcgtacctc 5280 gtgcgcctgg ggcagcgcgt tatccagcgc gagcacaatg caccgatcat cctcactggc 5340 tactcagcgc tgaataagct tctgggtcgc gaagtctaca ccagccacga acagctgggc 5400 ggtacgaaga tcatgtatcc gaacggggtt acccatctgc gcgtatccaa tgactacgag 5460 ggtgttcggg caattctgga ctggctcgct tttgtgcctc gtatgcaagg ggaacgacct 5520 ccgatgatcg actccatcga tgctgtggta cgcgaagtcg actacgatcc acgagtggct 5580 aacgaggata ttcgatacgc cattgaaggc aagtgggtcg gacccttcgc tcccgacggg 5640 gcaagtgtgg tctcgtcgtc gggctcgtcg gtgtcgtcca atggtccggt tgcgacaggt 5700 gcgagcgctg gactgccgcc cgtcaacatg gagagtgtcg gcggtgaagt tcggtatcgt 5760 tcgcggtcgc ggacggcttc gtatgcgggt gcgaatgcag caacagcagc agcaccagga 5820. gcaagtagct tggcgacctc tggagcggca cttgcgaatg ctacgggacg agcagcggct 5880 cgggtgagcc tcgtatcgac caagcctgca accggtgtcg aggaggttcc cgcggcgttg 5940 ccttcgggtt cgagttcact ggcgctccgc tcgaccgctg tcgctgcgcc tgtacctcag 6000 gagagagcaa agacggatgc ggatgcggat gcggatgcgg atgcggatgc cttggaggcc 6060 actggaactg cccgaggcgc agataccagc ggtgctgggg cgcacgcaag ccctgacacc cggcagccgc ccacactgga tgccctgaag actgcgaggt ttgcaccagc gcccgctgca 6180. ttgccgacga caggtttacg ctcgccacca cagtccccgg gttctgtttc tgcgtatagc 6240 ccgttggaga gtggctctcc attggagccg tcccttgacg aggatcacat cgctttcacg 6300 gacaccgatc ggatgcggat cgggtcgcct ttggtacacg cgaccgacgc ctgggatacg 6360 agcaacagcg ttcactcgag tagcgcgacg acgatggctg caggtgccgt ccggggctcg cgactgttca ctgcgtcatc gagtacgtcg gcggctgcgg gagcaagcgc cagcaatagc 6480 agcaacaacg gtagcagcaa tgcgaatacg aataccatga acaatgctgg gagtagcgtg gcatcggcag gcgcaccgct ggtagcatca attack 6600. gcatcggcag gcgcaccgct gccaatacca acaatatctt gggtagcggt ggtgttggtg cgacagcttc gctgaccgct 6720. tggagcggta accttccgcc gctaccgtac gttccaaata cgctcatggt ggtggacaac caagggtacg tgttcctggc aggcctcttc gaccgtcact cgtttcgaga gacgctcgct 6780 ggctgggcga agtccgttgt ggttggtaga gcacgccttg gtggtatccc cgtgggcgtt 6840 attgcgac agacggtgac cacagagac gttattcccc cggatccagc ggcgccagac tcgcgtgagc ttcgtgaaca gcaggccggt caagtctggt atccagattc atccttcaag actgcccaaa gtattcgtga ttttgatcgc gagggtttgc ctctgtttgt tctggccagc tggcgcggtt tctctggcgg agcacacgac atgttccagg aaatactcaa gtttggttca gagatcgttg acgcgcttcg cgagtactcg aaacccgtct ttgtgtacat accaccggga 7140 ggcgagcttc gaggcggtgc ctgggtcgtc ctcgataccg ccatcaatcc ccgctttatc 7200 gaaatgtacg ccgatgaaag tgcacgtggt ggtgtgctcg aacctgccgg taccgtggat 7260 atcaagttcc gcaccagaga tttgctgaag accatgcagc gcctgctacc atcgtctaga 7320 cgtgatgaga gcgattcgag tgcgtccgac gttgtcggca tgctgtcttt ggacgcatca 7380 tcatcatcag cagcagcaac agcagcaaca acaacaacaa caacaacaac aacaacaaca 7440 gcaggtaccg ctgggacgtc ctcttcgact gctgggcgta gcgagtcgcg ggtcacaaag 7500 tcaacggcgg acggcttgcc agccttcgag gcagacggtc ggagttcgct cgagcaaaca 7560 ctcttgccca tctttcagca gatagccatg acgtttgcgg atctgcacga tacagcgggt 7620 cggatgcaac ataagcgcgc gattcgccga gtggtaccct ggcgaacaag tcgcacgttt 7680 ttctactggc gccttcgacg tcggctcgcc gaggaggaac tgcggagtcg cgtttcacaa 7740 gcggatccgc aactcagcga cgcggagatc gatgcgctgc tgcggaaatg ggcaagagca 7800 catgaccccg ctctggacgg tgatatttac gaacttgatg atcctcgtgt cgtgcaatgg 7860 ctcgaggacg aattggatag ccagttggag cggcgtttgc acaaactccg cgaagcacgc 7920 gccacctggc aagctgtgga actggggaat acagcacccg aggcgttgct tgcggcaatt 7980 gagcgtattc tcgtacaaat ggatgaagtt ggtcgcaacg aatggttacg aacgcttagt 8040 gcgcgactcg agaacacctc atccggtttg ggtgatctcg aggcgcccag ctcgggtgag 8100 caggccgcac cacgacgtgt cttgtcgggt cgtggtctgc tgcgacgctt cctcggg 8157 <210> 20 <211> 2719 <212> PRT <213> Cyanidioschyzon merolae (a red alga) <400> 20 Met Gly Asp Leu Gly Glu His Ala Pro Ser Asn Gly Ser Ala Lys Asp 1 5 10 15 Ser Val Arg Arg Tyr Val Glu Glu His Gly Gly Ser Arg Pro Ile Gln 20 25 30 Arg Leu Leu Val Ala Asn Asn Gly Ile Ala Ala Val Lys Cys Ile Arg 35 40 45 Ser Met Arg Arg Trp Ala Tyr Glu Ala Phe Gly Ser Glu Arg Ala Leu 50 55 60 Glu Phe Val Ala Met Ala Thr Pro Glu Asp Val Gln Ala Asn Ala Asp 65 70 75 80 Tyr Ile Arg Leu Ala Asp Leu Phe Val Pro Val Pro Gly Gly Ser Asn 85 90 95 Asn His Asn Tyr Ala Asn Val Glu Leu Ile Val Asp Ile Ala Glu Arg 100 105 110 Asn Glu Cys Asp Ala Val Trp Ala Gly Trp Gly His Ala Ser Glu Asn 115 120 125 Pro Leu Leu Pro Ala Arg Leu Ala Glu Thr Gly Ile Ala Phe Leu Gly 130 135 140 Pro Asp Ala Ile Ala Met Arg Ala Leu Gly Asp Lys Ile Ser Ser Thr 145 150 155 160 Leu Leu Ala Gln Ser Ala Asp Val Pro Val Val Ser Trp Asn Gly Asp 165 170 175 Asp Leu Lys Val Thr Phe His Arg Glu Arg Gly Gly Ile Asp Glu Glu 180 185 190 Arg Tyr Arg Arg Ala Cys Val Ala Asn Val Asp Glu Ala Arg Ala Ala 195 200 205 Ala Asp Arg Ile Gly Tyr Pro Val Met Ile Lys Ala Ser Glu Gly Gly 210 215 220 Gly Gly Lys Gly Ile Arg Leu Cys Arg Arg Pro Glu Asp Val Arg Asp 225 230 235 240 Ala Phe Arg Gln Val Ala Gly Glu Ile Pro Gly Ser Pro Ile Phe Ile 245 250 255 Met Lys Met Val Asp Gln Ala Arg His Leu Glu Val Gln Ile Val Ala 260 265 270 Asp Glu Tyr Gly His Ala Ile Ala Leu Tyr Gly Arg Asp Cys Ser Val 275 280 285 Gln Arg Arg His Gln Lys Ile Ile Glu Glu Gly Pro Val Thr Ala Thr 290 295 300 Pro Pro Gln Val Trp Lys Glu Leu Glu His Gly Ala Val Arg Leu Ala 305 310 315 320 Lys Met Val Gly Tyr Val Gly Ala Gly Thr Val Glu Tyr Leu Tyr Asp 325 330 335 Gly Arg Arg Phe Tyr Phe Leu Glu Leu Asn Pro Arg Leu Gln Val Glu 340 345 350 His Pro Val Thr Glu Trp Ile Thr Gly Val Asn Leu Pro Ala Val Gln 355 360 365 Leu Gln Ile Ala Met Gly Ile Pro Leu Gln Arg Ile Ala Ala Ile Gln 370 375 380 Ala Leu Tyr Gly Asn Arg Pro Asp Leu Asp Leu Glu Arg His Gln Pro 385 390 395 400 Asn Pro Pro His Gly His Val Ile Ala Cys Arg Val Thr Ala Glu Asn 405 410 415 Pro Glu Glu Gly Phe Gln Pro Thr Ser Gly Ser Ile Gln Glu Leu Ser 420 425 430 Phe Arg Asn Thr Pro Asn Val Trp Gly Tyr Phe Ser Val Gly Ala Trp 435 440 445 Gly Gly Val His Glu Tyr Ala Asp Ser Gln Phe Gly His Leu Phe Ala 450 455 460 Trp Gly Ala Asp Arg Glu Met Ala Arg Arg Asn Met Val Leu Ala Leu 465 470 475 480 Lys Glu Leu Ser Ile Arg Gly Asp Ile Arg Thr Thr Val Glu Tyr Leu 485 490 495 Ile Thr Leu Leu Glu Leu Glu Ser Tyr Arg Glu Asn Arg Ile His Thr 500 505 510 Arg Trp Leu Asp Asn Leu Ile Ala Ser Lys Val Lys Pro Glu Arg Pro 515 520 525 Pro Phe His Ile Ala Val Val Leu Gly Ala Val His Gln Ala Tyr Arg 530 535 540 Ala Trp Ser Glu Arg Arg Arg Ser Phe Val Glu Ser Leu Ser Arg Gly 545 550 555 560 Gln Val Pro Gln Arg Ile Asp Ala Thr Phe Ile Glu Phe Gln Phe Glu 565 570 575 Leu Ile Tyr Asp Glu Leu Lys Tyr Ala Leu Ile Val Arg Gln Ala Gly 580 585 590 Pro Asn Ala Phe His Val Ser Leu Ala Gln His Pro Glu Glu Arg Val 595 600 605 Arg Val Asp Phe Arg Pro Leu Leu Asp Gly Gly Leu Leu Ile Met Cys 610 615 620 Asp Gly Arg Ser Phe Asn Thr His Ser Lys Glu Asp Ser Thr Gly Met 625 630 635 640 Arg Leu Ser Ile Asp Gly Arg Thr Cys Val Phe Pro Gly Glu Met Asp 645 650 655 Pro Thr Arg Ile Ala Ala Gln Ala Ser Gly Arg Leu Val Arg Tyr Leu 660 665 670 Val Ala Asn Glu Glu His Val Asp Gln Gly Gln Val Ile Ala Glu Ile 675 680 685 Glu Val Met Lys Met Tyr Leu Ser Val Gln Ala Pro Glu Pro Gly Thr 690 695 700 Ile His Leu Leu Lys Pro Ala Gly Ala Ala Leu Glu Pro Gly Glu Val 705 710 715 720 Phe Ala Gln Leu Asp Leu Asp Asp Pro Ser Lys Val Arg Arg Val Thr 725 730 735 Pro Phe Thr Gly Arg Phe Pro Thr Met Leu Pro Pro Gln Arg Leu Gly 740 745 750 Lys Lys Pro His Gln Arg Phe Glu Ser Ala Lys Gln Gln Ile Glu Ala 755 760 765 Leu Leu Asp Gly Tyr Asp Val Asp Met Glu Pro Leu Gly Met Leu Leu 770 775 780 Gln Leu Ala Ser Thr Glu Pro Ala Val Pro Ala Gly Lys Leu Gln Glu 785 790 795 800 Ala Leu Ser Phe Leu Ala Gly Arg Ile Pro Ser Thr Leu His Ala Ala 805 810 815 Leu Leu Gly Gln Val Phe Glu Leu Arg Ser Val Ala Arg Asp Asp His 820 825 830 Glu Arg Leu Gln Gln His Phe Arg Glu Met Lys Arg Met Ile Ser Asp 835 840 845 Phe Cys Glu Gln Leu Ala Ile Ala Ala Asp Arg Glu Ala Leu Glu Ala 850 855 860 Thr Leu Arg Pro Leu Thr Ser Ile Leu Glu Ala Tyr Val Arg Gly Gly 865 870 875 880 Leu Arg Gly Tyr His Glu His Leu Val Val Cys Leu Met Gln Arg Tyr 885 890 895 Val Asn Thr Glu Lys Tyr Phe Ala Gln Gly Arg Arg Leu Asp Glu Val 900 905 910 Leu Phe Asp Leu Arg Glu Gln His Arg Glu His Leu Glu Val Val Ala 915 920 925 Asp Ile Met Leu Ala His Ala Gln Leu Ala Arg Lys Asn Gln Leu Met 930 935 940 Leu Ala Leu Leu Asp His Ile Ala Ala Asp Ala Ser Leu Leu Arg Ser 945 950 955 960 Val Ile Val Arg Gln Cys Leu His Glu Val Ala Ala Phe Ile His Pro 965 970 975 Asp Tyr Ser Gln Leu Ala Leu Arg Ala Arg Leu Leu Leu Ala Ser Ser 980 985 990 Arg Arg Arg Ala Leu Pro Glu Arg Arg Glu Arg Leu Cys Lys Gln Ile 995 1000 1005 Glu Gln Ala Val His Ala Pro Thr Glu Glu Gln Cys Tyr Arg Leu 1010 1015 1020 Leu His Ala Val Val Glu Ser Gln Glu Ser Ile Leu Asp Ala Leu 1025 1030 1035 Val Ser Leu Thr Met Asn Pro Gly Val Pro Val Glu Ile Arg Lys 1040 1045 1050 Ala Ala Val Gln Cys Gln Ile Leu Arg Ala Tyr Lys Ala Tyr His 1055 1060 1065 Val Tyr Asp Leu Val Val Glu Leu Asp Glu Glu Leu Gly Phe Leu 1070 1075 1080 Arg Ala Leu Trp Arg Phe Gln Tyr Arg Ser Asn Leu Asn Ala Phe 1085 1090 1095 Gly Ser His Ser Ser Met Arg Ser Phe Pro Ala Ala Leu Val Ala 1100 1105 1110 Ala Ser Ala Pro Leu Ala Arg Arg Gln Leu Arg Ser Tyr Asp Ser 1115 1120 1125 Ala Asp Asn Leu Gln Asn Trp Gly Phe Arg Val Gly Met Leu Val 1130 1135 1140 Val Phe Glu Thr Leu Arg Ala Met Val Gln Gly Phe Asp Arg Val 1145 1150 1155 Leu Gln Val Phe Arg Ala Glu Thr Gly Gly Glu Thr Leu Ala Ser 1160 1165 1170 Pro Arg Thr Lys Ser Ser Ala Ala Val Asp Ser Ser Ala Asp Ser 1175 1180 1185 Ser Ala Glu Gly Ile Gly Val Asn Val Leu Thr Ile Ala Ile Pro 1190 1195 1200 Trp Thr Ala Val Glu Val Ala Asp Phe Ala Lys Tyr Leu Gly Val 1205 1210 1215 Ala Asp Ala Asp Asp Arg Leu Gln Ala Ser Asn Ala Asn Asp Ser 1220 1225 1230 Glu Gln Val Val Ala Leu Leu Thr Arg Phe Cys Arg Ser Ser Ala 1235 1240 1245 Glu Arg Lys Ala Ser Met Arg Ala Ala Gly Ile Lys Leu Val Thr 1250 1255 1260 Phe Leu Val Ala Pro Gly Glu Leu Cys Gln Arg Thr Leu Gly Ser 1265 1270 1275 Ser Leu Pro Arg Glu Ser Thr Tyr Pro Gly Phe Tyr Thr Leu Arg 1280 1285 1290 Ala Ser Leu Glu Tyr Ala Glu Asp Pro Ile Tyr Arg His Ile Asp 1295 1300 1305 Pro Pro Ala Ala Phe Gln Leu Glu Leu Asn Arg Leu Ala Asn Phe 1310 1315 1320 Arg Ile Thr Arg Phe Glu His Pro Asn Arg Ser Ile His Val Phe 1325 1330 1335 Tyr Ala Glu Asp Arg Thr Asp Lys Gly Asp Ala Arg Phe Phe Val 1340 1345 1350 Arg Ala Phe Val Arg Gln Ala Glu Val Tyr Ala Ser Pro Ser Asp 1355 1360 1365 Thr Ala Ala Val Ser Ile Pro Glu Ala Glu Arg Thr Phe Val Ala 1370 1375 1380 Cys Leu Asp Ala Leu Glu Thr Ala Arg Cys Asp Arg Arg Phe Arg 1385 1390 1395 Arg Thr Asp Phe Asn His Leu Phe Leu His Leu Ile Pro Arg Val 1400 1405 1410 Ser Ile Asp Val Asp Asp Val Glu Ala Ile Cys Thr Arg Leu Phe 1415 1420 1425 Tyr Arg Phe Ser Ser Arg Cys Trp Arg Leu Arg Val Phe Met Val 1430 1435 1440 Glu Ile Lys Val His Val Glu Lys Met Gly Pro Lys Pro Leu Arg 1445 1450 1455 Phe Ile Leu Tyr Asn Pro Thr Gly His Ser Leu Arg Val Glu Gly 1460 1465 1470 Tyr Val Glu Gln Gly Asp Arg Leu Leu Ser Leu Asp Ser Ser Asp 1475 1480 1485 Pro Gly His Leu His Gly Thr Pro Val Asp Glu Pro Tyr Gln Val 1490 1495 1500 Leu Asn Arg Ile Gln Arg Arg Arg Val Val Ala Gln Thr Leu Glu 1505 1510 1515 Thr Thr Tyr Val Tyr Asp Tyr Gln Glu Leu Phe Thr Lys Ala Leu 1520 1525 1530 His Glu Gln Trp Arg Arg Tyr Ser Gln Glu Arg Leu Leu Gly Gly 1535 1540 1545 Phe Arg Arg His Lys Ile Pro Leu Lys Leu Leu Thr Cys Thr Glu 1550 1555 1560 Leu Val Leu Gln Asp Glu Asp His Asp Glu Ser Glu Leu Ile Glu 1565 1570 1575 Thr Asn Arg Pro Ala Gly Glu Asn Glu Ile Gly Met Val Ala Trp 1580 1585 1590 Arg Tyr Thr Phe Phe Thr Pro Glu Tyr Pro Gln Gly Arg Asp Ala 1595 1600 1605 Ile Val Ile Ala Asn Asp Ile Thr Tyr Leu Ser Gly Ser Phe Gly 1610 1615 1620 Pro Arg Glu Asp Arg Leu Phe Ala Lys Ala Ser Gln Gln Ala Arg 1625 1630 1635 Lys Leu Gly Ile Pro Arg Ile Tyr Ile Ala Ala Asn Ser Gly Ala 1640 1645 1650 Arg Ile Gly Leu Ala Glu Glu Leu Arg Asn Val Phe Gln Val Gln 1655 1660 1665 Trp Lys Asp Ala His Asp Pro Ser Arg Gly Phe Asp Tyr Leu Tyr 1670 1675 1680 Leu Gly Leu Ala Asp Lys Leu Lys Tyr Glu Asp Glu Leu Gly Ile 1685 1690 1695 Val Arg Thr Arg Asp Val Gly Gly Gly Lys Tyr Ala Leu Val Asp 1700 1705 1710 Ile Tyr Gly Ala Glu Asp Gly Ile Gly Val Glu Asn Leu Met Gly 1715 1720 1725 Ser Ala Cys Ile Ala Ala Glu Thr Ser Ala Ala Tyr Asn Asp Cys 1730 1735 1740 Phe Thr Ile Thr Tyr Val Ala Ala Arg Cys Val Gly Ile Gly Ala 1745 1750 1755 Tyr Leu Val Arg Leu Gly Gln Arg Val Ile Gln Arg Glu His Asn 1760 1765 1770 Ala Pro Ile Ile Leu Thr Gly Tyr Ser Ala Leu Asn Lys Leu Leu 1775 1780 1785 Gly Arg Glu Val Tyr Thr Ser His Glu Gln Leu Gly Gly Thr Lys 1790 1795 1800 Ile Met Tyr Pro Asn Gly Val Thr His Leu Arg Val Ser Asn Asp 1805 1810 1815 Tyr Glu Gly Val Arg Ala Ile Leu Asp Trp Leu Ala Phe Val Pro 1820 1825 1830 Arg Met Gln Gly Glu Arg Pro Pro Met Ile Asp Ser Ile Asp Ala 1835 1840 1845 Val Val Arg Glu Val Asp Tyr Asp Pro Arg Val Ala Asn Glu Asp 1850 1855 1860 Ile Arg Tyr Ala Ile Glu Gly Lys Trp Val Gly Pro Phe Ala Pro 1865 1870 1875 Asp Gly Ala Ser Val Val Ser Ser Ser Gly Ser Ser Val Ser Ser 1880 1885 1890 Asn Gly Pro Val Ala Thr Gly Ala Ser Ala Gly Leu Pro Pro Val 1895 1900 1905 Asn Met Glu Ser Val Gly Gly Glu Val Arg Tyr Arg Ser Arg Ser 1910 1915 1920 Arg Thr Ala Ser Tyr Ala Gly Ala Asn Ala Ala Thr Ala Ala Ala 1925 1930 1935 Pro Gly Ala Ser Ser Leu Ala Thr Ser Gly Ala Ala Leu Ala Asn 1940 1945 1950 Ala Thr Gly Arg Ala Ala Ala Arg Val Ser Leu Val Ser Thr Lys 1955 1960 1965 Pro Ala Thr Gly Val Glu Glu Val Pro Ala Ala Leu Pro Ser Gly 1970 1975 1980 Ser Ser Ser Leu Ala Leu Arg Ser Thr Ala Val Ala Ala Pro Val 1985 1990 1995 Pro Gln Glu Arg Ala Lys Thr Asp Ala Asp Ala Asp Ala Asp Ala 2000 2005 2010 Asp Ala Asp Ala Leu Glu Ala Thr Gly Thr Ala Arg Gly Ala Asp 2015 2020 2025 Thr Ser Gly Ala Gly Ala His Ala Ser Pro Asp Thr Arg Gln Pro 2030 2035 2040 Pro Thr Leu Asp Ala Leu Lys Thr Ala Arg Phe Ala Pro Ala Pro 2045 2050 2055 Ala Ala Leu Pro Thr Thr Gly Leu Arg Ser Pro Pro Gln Ser Pro 2060 2065 2070 Gly Ser Val Ser Ala Tyr Ser Pro Leu Glu Ser Gly Ser Pro Leu 2075 2080 2085 Glu Pro Ser Leu Asp Glu Asp His Ile Ala Phe Thr Asp Thr Asp 2090 2095 2100 Arg Met Arg Ile Gly Ser Pro Leu Val His Ala Thr Asp Ala Trp 2105 2110 2115 Asp Thr Ser Asn Ser Val His Ser Ser Ser Ala Thr Thr Met Ala 2120 2125 2130 Ala Gly Ala Val Arg Gly Ser Arg Leu Phe Thr Ala Ser Ser Ser 2135 2140 2145 Thr Ser Ala Ala Ala Gly Ala Ser Ala Ser Asn Ser Ser Asn Asn 2150 2155 2160 Gly Ser Ser Asn Ala Asn Thr Asn Thr Met Asn Asn Ala Gly Ser 2165 2170 2175 Ser Val Ala Ser Ala Gly Ala Pro Leu Asn Thr Thr Gly Ser Ile 2180 2185 2190 Asn Thr Asn Thr Asn Ala Asn Ala Asn Thr Asn Asn Ile Leu Gly 2195 2200 2205 Ser Gly Gly Val Gly Ala Thr Ala Ser Leu Thr Ala Trp Ser Gly 2210 2215 2220 Asn Leu Pro Pro Leu Pro Tyr Val Pro Asn Thr Leu Met Val Val 2225 2230 2235 Asp Asn Gln Gly Tyr Val Phe Leu Ala Gly Leu Phe Asp Arg His 2240 2245 2250 Ser Phe Arg Glu Thr Leu Ala Gly Trp Ala Lys Ser Val Val Val 2255 2260 2265 Gly Arg Ala Arg Leu Gly Gly Ile Pro Val Gly Val Ile Ala Thr 2270 2275 2280 Gln Thr Val Thr Thr Glu Lys Val Ile Pro Pro Asp Pro Ala Ala 2285 2290 2295 Pro Asp Ser Arg Glu Leu Arg Glu Gln Gln Ala Gly Gln Val Trp 2300 2305 2310 Tyr Pro Asp Ser Ser Phe Lys Thr Ala Gln Ser Ile Arg Asp Phe 2315 2320 2325 Asp Arg Glu Gly Leu Pro Leu Phe Val Leu Ala Ser Trp Arg Gly 2330 2335 2340 Phe Ser Gly Gly Ala His Asp Met Phe Gln Glu Ile Leu Lys Phe 2345 2350 2355 Gly Ser Glu Ile Val Asp Ala Leu Arg Glu Tyr Ser Lys Pro Val 2360 2365 2370 Phe Val Tyr Ile Pro Pro Gly Gly Glu Leu Arg Gly Gly Ala Trp 2375 2380 2385 Val Val Leu Asp Thr Ala Ile Asn Pro Arg Phe Ile Glu Met Tyr 2390 2395 2400 Ala Asp Glu Ser Ala Arg Gly Gly Val Leu Glu Pro Ala Gly Thr 2405 2410 2415 Val Asp Ile Lys Phe Arg Thr Arg Asp Leu Leu Lys Thr Met Gln 2420 2425 2430 Arg Leu Leu Pro Ser Ser Arg Arg Asp Glu Ser Asp Ser Ser Ala 2435 2440 2445 Ser Asp Val Val Gly Met Leu Ser Leu Asp Ala Ser Ser Ser Ser 2450 2455 2460 Ala Ala Ala Thr Ala Ala Thr Thr Thr Thr Thr Thr Thr Thr Thr 2465 2470 2475 Thr Thr Ala Gly Thr Ala Gly Thr Ser Ser Ser Thr Ala Gly Arg 2480 2485 2490 Ser Glu Ser Arg Val Thr Lys Ser Thr Ala Asp Gly Leu Pro Ala 2495 2500 2505 Phe Glu Ala Asp Gly Arg Ser Ser Leu Glu Gln Thr Leu Leu Pro 2510 2515 2520 Ile Phe Gln Gln Ile Ala Met Thr Phe Ala Asp Leu His Asp Thr 2525 2530 2535 Ala Gly Arg Met Gln His Lys Arg Ala Ile Arg Arg Val Val Pro 2540 2545 2550 Trp Arg Thr Ser Arg Thr Phe Phe Tyr Trp Arg Leu Arg Arg Arg 2555 2560 2565 Leu Ala Glu Glu Glu Leu Arg Ser Arg Val Ser Gln Ala Asp Pro 2570 2575 2580 Gln Leu Ser Asp Ala Glu Ile Asp Ala Leu Leu Arg Lys Trp Ala 2585 ​​2590 2595 Arg Ala His Asp Pro Ala Leu Asp Gly Asp Ile Tyr Glu Leu Asp 2600 2605 2610 Asp Pro Arg Val Val Gln Trp Leu Glu Asp Glu Leu Asp Ser Gln 2615 2620 2625 Leu Glu Arg Arg Leu His Lys Leu Arg Glu Ala Arg Ala Thr Trp 2630 2635 2640 Gln Ala Val Glu Leu Gly Asn Thr Ala Pro Glu Ala Leu Leu Ala 2645 2650 2655 Ala Ile Glu Arg Ile Leu Val Gln Met Asp Glu Val Gly Arg Asn 2660 2665 2670 Glu Trp Leu Arg Thr Leu Ser Ala Arg Leu Glu Asn Thr Ser Ser 2675 2680 2685 Gly Leu Gly Asp Leu Glu Ala Pro Ser Ser Gly Glu Gln Ala Ala 2690 2695 2700 Pro Arg Arg Val Leu Ser Gly Arg Gly Leu Leu Arg Arg Phe Leu 2705 2710 2715 Gly <210> 21 <211> 1596 <212> DNA <213> Cyanidioschyzon merolae (a red alga) <400> 21 atgcttctgg cggaaagcct cggcggcgtc gcttttgtca ccgttcacac ggtgctgtgc 60 gtcgctgtac tgtacttacc gtggaaccgc ggcgaccgag aaagcttccg tgtcacttgt 120 ctaggggctt tgagcacttg cgctgttatc gctctgagcc tggtgaatct tcagttgttc 180 ttcactggct tgacctggcg ggagctgtcg cggacgtatc actggctgag gccgtggcac 240 agctggcaag tctatctact ggactctggc ggttttttgg tgctcctttt tctgcacgag 300 tacagaaaaa gacagcagac tccagttaag agttcgtcat cgagcacgtg tgcatcgacg 360 tcgcagtccg cttgcccgaa tgcggagaac ggcatctatc gaaacgagag gcgaaatacc 420 tggtgcatgt tccgaagcct gctgatcgca ccgataacag aggagctcct cttccgttgt 480 gtattcgatg cagcaatgag atcagctcaa gtccctgaac ttgccagcat gattttcaac 540 ggagtcatgt ttgccgtagc gcacgcgcat cactatttcc ggcaccagag caggtcactt 600 ctcggaaagc agcttcttgt aacgttttgc tttggttgcg tccaagtcgt ttgcctgaga 660 cggacggatt actccctgtg ggcctgcatc gcaacgcacg ctttggcgaa cgcgctcgat 720 ctgcagaaag ttttcagtga taggggagcg tcacattttc tctacggcga cgtaggccat 780 cagttaggag ccctgctgca agctgcagca ccgctcgtat tcatcttgcg ctatgcgctt 840 gacatggcca tgcaagcgag tccgttccgg tcctcgatcc tccaggtgca tgcggacccg 900 gggctgctcg aggaacggcg tgaaatcatg cgtccaacga cgccgttttc gctcttcttg 960 atagaatatc atcagcgtga tccgatcttc gggcgcttca tggctctctg ctccatgctg 1020 cctcaacttt tattcgccgc agaggtcaca gctgtctact gttggcgaag tccgcgcgca 1080 ttgctcctag ctgttggcca gattgtgaac gagacgctga gctacgcact caagcgatcg 1140 tgtcggatac ctcgcccgcc aatcgctcga ctggaagcag atgcgttcgg ttggccctct 1200 tcacacgctc agttcatgtc ctacttgtac gttttctacg tactctatgt gtctaggcca 1260 cagcgcaagg catcatcgag ccacaacggt gaaatgatgc accagacgct tccgaagcgg 1320 aggaagacga ccgattcaat ttcggaaacc gtggcagtga tgtttttgct aggtctgtca 1380 tccgttctgg tggctgcctc aagagtatac ctggcatatc attatcccag tcaggtttgg 1440 tatggaatca tcatgggcac tattttcgcg ataacctggt tccttgcgag tgaaaacgtt 1500 ttcttatcct acgttcgatc attgcggatt ctcgtatggc tcgggttcgg cgaagatgac 1560 tctgacctgt ttcgcagtat actcagagat gtgcat 1596 <210> 22 <211> 532 <212> PRT <213> Cyanidioschyzon merolae (a red alga) <400> 22 Met Leu Leu Ala Glu Ser Leu Gly Gly Val Ala Phe Val Thr Val His 1 5 10 15 Thr Val Leu Cys Val Ala Val Leu Tyr Leu Pro Trp Asn Arg Gly Asp 20 25 30 Arg Glu Ser Phe Arg Val Thr Cys Leu Gly Ala Leu Ser Thr Cys Ala 35 40 45 Val Ile Ala Leu Ser Leu Val Asn Leu Gln Leu Phe Phe Thr Gly Leu 50 55 60 Thr Trp Arg Glu Leu Ser Arg Thr Tyr His Trp Leu Arg Pro Trp His 65 70 75 80 Ser Trp Gln Val Tyr Leu Leu Asp Ser Gly Gly Phe Leu Val Leu Leu 85 90 95 Phe Leu His Glu Tyr Arg Lys Arg Gln Gln Thr Pro Val Lys Ser Ser 100 105 110 Ser Ser Ser Thr Cys Ala Ser Thr Ser Gln Ser Ala Cys Pro Asn Ala 115 120 125 Glu Asn Gly Ile Tyr Arg Asn Glu Arg Arg Asn Thr Trp Cys Met Phe 130 135 140 Arg Ser Leu Leu Ile Ala Pro Ile Thr Glu Glu Leu Leu Phe Arg Cys 145 150 155 160 Val Phe Asp Ala Ala Met Arg Ser Ala Gln Val Pro Glu Leu Ala Ser 165 170 175 Met Ile Phe Asn Gly Val Met Phe Ala Val Ala His Ala His His Tyr 180 185 190 Phe Arg His Gln Ser Arg Ser Leu Leu Gly Lys Gln Leu Leu Val Thr 195 200 205 Phe Cys Phe Gly Cys Val Gln Val Val Cys Leu Arg Arg Thr Asp Tyr 210 215 220 Ser Leu Trp Ala Cys Ile Ala Thr His Ala Leu Ala Asn Ala Leu Asp 225 230 235 240 Leu Gln Lys Val Phe Ser Asp Arg Gly Ala Ser His Phe Leu Tyr Gly 245 250 255 Asp Val Gly His Gln Leu Gly Ala Leu Leu Gln Ala Ala Ala Pro Leu 260 265 270 Val Phe Ile Leu Arg Tyr Ala Leu Asp Met Ala Met Gln Ala Ser Pro 275 280 285 Phe Arg Ser Ser Ile Leu Gln Val His Ala Asp Pro Gly Leu Leu Glu 290 295 300 Glu Arg Arg Glu Ile Met Arg Pro Thr Thr Pro Phe Ser Leu Phe Leu 305 310 315 320 Ile Glu Tyr His Gln Arg Asp Pro Ile Phe Gly Arg Phe Met Ala Leu 325 330 335 Cys Ser Met Leu Pro Gln Leu Leu Phe Ala Ala Glu Val Thr Ala Val 340 345 350 Tyr Cys Trp Arg Ser Pro Arg Ala Leu Leu Leu Ala Val Gly Gln Ile 355 360 365 Val Asn Glu Thr Leu Ser Tyr Ala Leu Lys Arg Ser Cys Arg Ile Pro 370 375 380 Arg Pro Pro Ile Ala Arg Leu Glu Ala Asp Ala Phe Gly Trp Pro Ser 385 390 395 400 Ser His Ala Gln Phe Met Ser Tyr Leu Tyr Val Phe Tyr Val Leu Tyr 405 410 415 Val Ser Arg Pro Gln Arg Lys Ala Ser Ser Ser His Asn Gly Glu Met 420 425 430 Met His Gln Thr Leu Pro Lys Arg Arg Lys Thr Thr Asp Ser Ile Ser 435 440 445 Glu Thr Val Ala Val Met Phe Leu Leu Gly Leu Ser Ser Val Leu Val 450 455 460 Ala Ala Ser Arg Val Tyr Leu Ala Tyr His Tyr Pro Ser Gln Val Trp 465 470 475 480 Tyr Gly Ile Ile Met Gly Thr Ile Phe Ala Ile Thr Trp Phe Leu Ala 485 490 495 Ser Glu Asn Val Phe Leu Ser Tyr Val Arg Ser Leu Arg Ile Leu Val 500 50​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​cgatggtttc tcgaacagtt ggctttagag ggcttcgtag tagtcgcgac gccgtaccga 420 ctttctttcg atcacctttg gacgatggat gacgtgctca ccaggttttc agcagcagcg 480 ggaatgctcg ctctcgatta cggtcccatc cctgttgttg gaatcggtca ctccctgggc 540 gctctcctgc acacccttgg aggcagcttg ttttgcaacg cggatgggta caaggctgcc 600 aacgtactga ttgcattcaa taatcgccgc gcggaagatg cgataccttt attcagagag 660 tttatcgcgc ccgtggttaa gacggttgcc caaactggtg aactctctga tgtgatcgag 720 cggctcgtga tcgatgggcc tgcgacgttt gacacgctgt tcgacactgt tacagatgtt 780 gtgtttcctg gttctcggga tagtgagctt ttcgccttgg tgcgacagtc aagggcgctt 840 gcgcagcaga ttccaccttt atttgctgag gtcgctgatg gagcttttgc attcaaccct 900 gatcctatcg aggtcatgga ggcaattcgc acactataca aagttcgtca aacgcttatt 960 gtgagcttca aaaacgacat cctcgatgac tcgcgttcgc ttcagaaagc actagaccct 1020 gagcgaggcg ctacagtaat tcgtctcggc ggaactcact taaccccctg cgcacaggat 1080 ttcctcgacc cacggctccg gcaaagcagc tttttctctt catttacgtg cgaaaaggat 1140 aacgacacat cctacatact ccgcgaagct atgcgcagaa ttgtgcttcg cgaagcaatg 1200 ctcatgaaaa acagtgttgt ggcgtattta gatcgcgctc tcggaatgga a 1251 <210> 24 <211> 417 <212> PRT <213> Cyanidioschyzon merolae (a red alga) <400> 24 Met Ser Cys Lys Gly Ala Ala Phe Gly Phe Ala Thr Leu Asn Gly Leu 1 5 10 15 Ser Val Arg Arg His Leu Phe Cys Arg Ser Ser Ala Asn Arg Arg Val 20 25 30 Cys Gly Lys Leu Phe Thr Gly Ser Gln Cys Pro Arg Ala Pro Arg Tyr 35 40 45 Phe Leu Val His Thr Asp Lys Ser Ala Gly Asn Thr Pro Ser Thr Arg 50 55 60 Gly Trp Glu Glu Tyr Ala Gly Asn Phe Ile Phe Arg Pro Pro His Val 65 70 75 80 Ile Val Arg Pro Ser Gln Arg Thr Ala Tyr Gly Thr Gly Asn Gly Thr 85 90 95 Pro Leu Pro Pro Lys Ala Leu Val His Phe Leu Gly Gly Ala Phe Val 100 105 110 Gly Ala Ala Pro Gln Val Ala Tyr Arg Trp Phe Leu Glu Gln Leu Ala 115 120 125 Leu Glu Gly Phe Val Val Val Ala Thr Pro Tyr Arg Leu Ser Phe Asp 130 135 140 His Leu Trp Thr Met Asp Asp Val Leu Thr Arg Phe Ser Ala Ala Ala 145 150 155 160 Gly Met Leu Ala Leu Asp Tyr Gly Pro Ile Pro Val Val Gly Ile Gly 165 170 175 His Ser Leu Gly Ala Leu Leu His Thr Leu Gly Gly Ser Leu Phe Cys 180 185 190 Asn Ala Asp Gly Tyr Lys Ala Ala Asn Val Leu Ile Ala Phe Asn Asn 195 200 205 Arg Arg Ala Glu Asp Ala Ile Pro Leu Phe Arg Glu Phe Ile Ala Pro 210 215 220 Val Val Lys Thr Val Ala Gln Thr Gly Glu Leu Ser Asp Val Ile Glu 225 230 235 240 Arg Leu Val Ile Asp Gly Pro Ala Thr Phe Asp Thr Leu Phe Asp Thr 245 250 255 Val Thr Asp Val Val Phe Pro Gly Ser Arg Asp Ser Glu Leu Phe Ala 260 265 270 Leu Val Arg Gln Ser Arg Ala Leu Ala Gln Gln Ile Pro Pro Leu Phe 275 280 285 Ala Glu Val Ala Asp Gly Ala Phe Ala Phe Asn Pro Asp Pro Ile Glu 290 295 300 Val Met Glu Ala Ile Arg Thr Leu Tyr Lys Val Arg Gln Thr Leu Ile 305 310 315 320 Val Ser Phe Lys Asn Asp Ile Leu Asp Asp Ser Arg Ser Leu Gln Lys 325 330 335 Ala Leu Asp Pro Glu Arg Gly Ala Thr Val Ile Arg Leu Gly Gly Thr 340 345 350 His Leu Thr Pro Cys Ala Gln Asp Phe Leu Asp Pro Arg Leu Arg Gln 355 360 365 Ser Ser Phe Phe Ser Ser Phe Thr Cys Glu Lys Asp Asn Asp Thr Ser 370 375 380 Tyr Ile Leu Arg Glu Ala Met Arg Arg Ile Val Leu Arg Glu Ala Met 385 390 395 400 Leu Met Lys Asn Ser Val Val Ala Tyr Leu Asp Arg Ala Leu Gly Met 405 410 415 Glu <210> 25 <211> 447 <212> DNA <213> Cyanidioschyzon merolae (a red alga) <400> 25 atgaacgact accaaaagat tggggttact ttgagcgcaa tttccgtttt attttatggg 60 ctcggtgttg tactcttttt tgatacgggg ctgatctcca tagcaagtgt tcttttcacc 120 tcgtcgctct ttttcatcct gggcttcaag agagccgccc gtttcttctt ctcaagaaga 180 aaacttcgcg cgagtgcgct cttcttcggc ggctttggcc tggtgctgtt gcgctggcca 240 gtgctgggga ctctggtcca agcggtcggt gcgctctggc tattcctgag cttcattccg 300 atcgcgatga cgtttcttcg acaggttccg gttctgggtc agcttgtcga caccagcatt 360 gttcggcgga tactgcgccg cctatcggca gcctctggat acctggaaac gaacggtggc 420 atcggtttcg aaccgaaact cccggta 447 <210> 26 <211> 149 <212> PRT <213> Cyanidioschyzon merolae (a red alga) <400> 26 Met Asn Asp Tyr Gln Lys Ile Gly Val Thr Leu Ser Ala Ile Ser Val 1 5 10 15 Leu Phe Tyr Gly Leu Gly Val Val Leu Phe Phe Asp Thr Gly Leu Ile 20 25 30 Ser Ile Ala Ser Val Leu Phe Thr Ser Ser Leu Phe Phe Ile Leu Gly 35 40 45 Phe Lys Arg Ala Ala Arg Phe Phe Phe Ser Arg Arg Lys Leu Arg Ala 50 55 60 Ser Ala Leu Phe Phe Gly Gly Phe Gly Leu Val Leu Leu Arg Trp Pro 65 70 75 80 Val Leu Gly Thr Leu Val Gln Ala Val Gly Ala Leu Trp Leu Phe Leu 85 90 95 Ser Phe Ile Pro Ile Ala Met Thr Phe Leu Arg Gln Val Pro Val Leu 100 105 110 Gly Gln Leu Val Asp Thr Ser Ile Val Arg Arg Ile Leu Arg Arg Leu 115 120 125 Ser Ala Ala Ser Gly Tyr Leu Glu Thr Asn Gly Gly Ile Gly Phe Glu 130 135 140 Pro Lys Leo Pro Val 145 <210> 27 <211> 2118 <212> Ms <213> Cyanidioschyzon merolae <400> 27 atgcatcagc aggaccctga cgagataccg ctttctgaac tccttgggaa ctctggagaa 60 tcggtgccga gagttgtttc agttgccaga caaggtggct tcggcgcacc gtcgatacag 120 gttgccaaga agtcttcctc agctgcactt tcggaagatg aactgcctct agctctactt 180 attggtacgg aaagaaagaa ggagtgcagt agaaggaagg gcaaagtcga accatgtcga 240 aaagcgccga aaaggaaatg cgaacgaatg aaagccgaaa agcaccgcaa ggcgaaaaag 300 ttgcaggcaa caagtactaa acatagagcg gaagatgaat cgatcaagtg gtgcagcctc 360 gaacataacg gagtcttatt ccctccggaa tatgaaccgc acggacggcc tctactctac 420 gatggctgcg aaatagcgct cccgcccgag gccgaagaag tggccacctt ttatgcgtcg 480 aagttgggaa ctgtttattt agaaaaggaa acatttcgga aaaacttttt tgacgacttt 540 cggaagacct tgcccgttga tttacggaag cgcatcgtga aactcgaaca ctgtgacttc 600 agccgtatcc gggagtatct cgacgagctg aaagaacgaa aacaaagcat gcctccaagc 660 aaacggaaag agttacgaga agcggaagcg caaagagttg cgcattacac ggttgcaatc 720 gttgatggac gcaaggagaa agtggccaat tatcgtgtcg agcctccggg tctctctcctg 780 gggcgcggtg accatccgct aatggggccga gtcaacgtc gcatattccc agaggatgtc 840 acactgaatc ttggtcgtga cgcacctatt ccgccgtgtc catttaagg ccatgactgg 900 ggatccatcg tgcacaacca gaggtcacg tggctggctt gctggcgaga tccaatctcc 960 gatgagtaca agtacgtctg gctgagtgcg tcttcgcact tcaggcgat gagcgatca 1020 gaaaaatttg agaaggcgca gcagctgagc aagcatatca cgaagatacg caatgagtac 1080 acgaaaggat tagaatctgc tgataggcac acgcagcaga gatccgtcgc actgtatctt 1140 attgataagc ttgcactccg agtcgggaac gagaaggg agacgaagc ggacactgtt 1200 ggttgctgct ccctgcggct agaacacata acccttcaag agcctatatat agtgcagctc 1260 gactttctcg gcaaggattc gatgcgatac ttacagag tgcgagtcga agcgctagtt 1320 tttcatcgcc tgcgagagtt ctgaaagga aaacgagtat cagataatgt attcgacgaa 1380 ctgaaagtcg aggaacttaa tgactttg aaagcctaa tgccggcct atctgcgaaa 1440 gtttttcgca cctacaatgc atcttacacc cttgataagc ttttgcacag cgtgaaaacc cctggaccgg atattcattc gcgccttctg ttctacaatg aggcgaacaa agatgtagct gtattatgta accatcagcg atcccttcca aagactcatt cgttaatgct tgaaagactg cgcgataagc tagaggagag cagagcgtat cttgaggcgc tcaaaatggc tcgaggaag 1680. gctcaggcat ctccagacag ccgcgcacaa gtaacacgct ggcgacggcc agtagttgag attcccgagg actgctccct cgctgagcga aagcgcattc gcgaggaggc cgaaaagcga ccgaaagaaa aacaggtagt gaacatgggg ctcgcgtcta tcacacgagg aatagcgcaa acacaggaa aaatcaggcg cctagaggct gacctgaaaa cgagggattc gctcgctacg gtgtcgttga gcacctctaa gatcaactat ttagatccgc gaataacagt tgcttggtgc aaaagacatg aggttcctat tgagcgaata ttccctcgcg cgctacagga aaagtttatg tggagtatgg gggtaagcga ggactttcgt ttcccaatca gcaacgtcgt ttcgaacgat ccctcagggg cttccaca 2118 <210> 28 <211> 706 <212> PRT <213> Cyanidioschyzon merolae (a red alga) <400> 28 Met His Gln Gln Asp Pro Asp Glu Ile Pro Leu Ser Glu Leu Leu Gly 1 5 10 15 Asn Ser Gly Glu Ser Val Pro Arg Val Val Ser Val Ala Arg Gln Gly 20 25 30 Gly Phe Gly Ala Pro Ser Ile Gln Val Ala Lys Lys Ser Ser Ser Ala 35 40 45 Ala Leu Ser Glu Asp Glu Leu Pro Leu Ala Leu Leu Ile Gly Thr Glu 50 55 60 Arg Lys Lys Glu Cys Ser Arg Arg Lys Gly Lys Val Glu Pro Cys Arg 65 70 75 80 Lys Ala Pro Lys Arg Lys Cys Glu Arg Met Lys Ala Glu Lys His Arg 85 90 95 Lys Ala Lys Lys Leu Gln Ala Thr Ser Thr Lys His Arg Ala Glu Asp 100 105 110 Glu Ser Ile Lys Trp Cys Ser Leu Glu His Asn Gly Val Leu Phe Pro 115 120 125 Pro Glu Tyr Glu Pro His Gly Arg Pro Leu Leu Tyr Asp Gly Cys Glu 130 135 140 Ile Ala Leu Pro Pro Glu Ala Glu Glu Val Ala Thr Phe Tyr Ala Ser 145 150 155 160 Lys Leu Gly Thr Val Tyr Leu Glu Lys Glu Thr Phe Arg Lys Asn Phe 165 170 175 Phe Asp Asp Phe Arg Lys Thr Leu Pro Val Asp Leu Arg Lys Arg Ile 180 185 190 Val Lys Leu Glu His Cys Asp Phe Ser Arg Ile Arg Glu Tyr Leu Asp 195 200 205 Glu Leu Lys Glu Arg Lys Gln Ser Met Pro Pro Ser Lys Arg Lys Glu 210 215 220 Leu Arg Glu Ala Glu Ala Gln Arg Val Ala His Tyr Thr Val Ala Ile 225 230 235 240 Val Asp Gly Arg Lys Glu Lys Val Ala Asn Tyr Arg Val Glu Pro Pro 245 250 255 Gly Leu Phe Leu Gly Arg Gly Asp His Pro Leu Met Gly Arg Val Lys 260 265 270 Arg Arg Ile Phe Pro Glu Asp Val Thr Leu Asn Leu Gly Arg Asp Ala 275 280 285 Pro Ile Pro Pro Cys Pro Phe Lys Gly His Asp Trp Gly Ser Ile Val 290 295 300 His Asn Gln Arg Val Thr Trp Leu Ala Cys Trp Arg Asp Pro Ile Ser 305 310 315 320 Asp Glu Tyr Lys Tyr Val Trp Leu Ser Ala Ser Ser His Phe Lys Ala 325 330 335 Met Ser Asp Gln Glu Lys Phe Glu Lys Ala Gln Gln Leu Ser Lys His 340 345 350 Ile Thr Lys Ile Arg Asn Glu Tyr Thr Lys Gly Leu Glu Ser Ala Asp 355 360 365 Arg His Thr Gln Gln Arg Ser Val Ala Leu Tyr Leu Ile Asp Lys Leu 370 375 380 Ala Leu Arg Val Gly Asn Glu Lys Gly Glu Asp Glu Ala Asp Thr Val 385 390 395 400 Gly Cys Cys Ser Leu Arg Val Glu His Ile Thr Leu Gln Glu Pro Asn 405 410 415 Ile Val Gln Leu Asp Phe Leu Gly Lys Asp Ser Met Arg Tyr Phe Asn 420 425 430 Arg Val Arg Val Glu Ala Leu Val Phe His Arg Leu Arg Glu Phe Leu 435 440 445 Lys Gly Lys Arg Val Ser Asp Asn Val Phe Asp Glu Leu Lys Val Glu 450 455 460 Glu Leu Asn Asp Tyr Leu Lys Ser Leu Met Pro Gly Leu Ser Ala Lys 465 470 475 480 Val Phe Arg Thr Tyr Asn Ala Ser Tyr Thr Leu Asp Lys Leu Leu His 485 490 495 Ser Val Lys Thr Pro Gly Pro Asp Ile His Ser Arg Leu Leu Phe Tyr 500 505 510 Asn Glu Ala Asn Lys Asp Val Ala Val Leu Cys Asn His Gln Arg Ser 515 520 525 Leu Pro Lys Thr His Ser Leu Met Leu Glu Arg Leu Arg Asp Lys Leu 530 535 540 Glu Glu Ser Arg Ala Tyr Leu Glu Ala Leu Lys Met Ala Arg Gly Lys 545 550 555 560 Ala Gln Ala Ser Pro Asp Ser Arg Ala Gln Val Thr Arg Trp Arg Arg 565 570 575 Pro Val Val Glu Ile Pro Glu Asp Cys Ser Leu Ala Glu Arg Lys Arg 580 585 590 Ile Arg Glu Glu Ala Glu Lys Arg Pro Lys Glu Lys Gln Val Val Asn 595 600 605 Met Gly Leu Ala Ser Ile Thr Arg Gly Ile Ala Gln Thr Gln Glu Lys 610 615 620 Ile Arg Arg Leu Glu Ala Asp Leu Lys Thr Arg Asp Ser Leu Ala Thr 625 630 635 640 Val Ser Leu Ser Thr Ser Lys Ile Asn Tyr Leu Asp Pro Arg Ile Thr 645 650 655 Val Ala Trp Cys Lys Arg His Glu Val Pro Ile Glu Arg Ile Phe Pro 660 665 670 Arg Ala Leu Gln Glu Lys Phe Met Trp Ser Met Gly Val Ser Glu Asp 675 680 685 Phe Arg Phe Pro Ile Ser Asn Val Val Ser Asn Asp Pro Ser Gly Ala 690 695 700 Ser Thr 705 <210> 29 <211> 600 <212> DNA <213> Cyanidioschyzon merolae (a red alga) <400> 29 cgacgagaac gtataaggag tgcgcacggc gttttgttac aataccgata gatgagtttc 60 gaacatcgca ttcacaccat gagcgggggc gcacgctcca gagagtggag atggaaaagt 120 gccagcggag ccctgaggat gcaaaaaagt acggaccgct gacggaagag caaatggaaa 180 ggagggcgaa acttcgaggg ctacttgcat tagtaagtac aaccaacgat ggtaagaaac 240 gtatggagtt tgcgaaccga gactttaacg ctgccatcaa tatcaggaga tgtgcggtgc 300 tggagacgag acctccagag tgaacaagaa ggtacttttt tggacaacct tctaaggtcg 360 aactatatga gataaaattg gaagaagtag ttggtggccg gtccaaaaag acggggaggc 420 gtctgcacat cagttggaga cgttttgtcc aaggcgcgcc gatcgccact actgtacacg 480 gccggcgaga acgtggcgag aatacgcaag cgagctcgcc ggctgcgctc gctgcaccac 540 cgtcttttga ctcacaactt cgcgatatct ttgttctctg tgtttcttcg ttcgttgacc 600 <210> 30 <211> 599 <212> DNA <213> Cyanidioschyzon merolae (a red alga) <400> 30 gcctcgaggg cctctctgcg tcgtcgccga agagctgaca gaaagcgttc caggtgcgcg 60 agcctcactt gggcatatac cagtgagtgg gtgcgggcac cactagtccg gtgaactcgg 120 ccggtcagcc aaatcctcca cccggaaggt tcgcaccgtc actgcagcga gcgcagtcca 180 agcttctagg cgcggtaggc gtgcaggatg cgcgtccaat tcggcaagat gcgctgccgg 240 catgcccgcc catgtgtcgg acccattccg tgcaagcgag gcacgtcgag cgatttgggg 300 cgcgtgcata ggcccgcgca cgatgaatga tctggttgca tgcgtacaca aacaggactt 360 ttctgaccat gatatcccta ttcggacatg acgcatcgcg cacttccgca acacctcgtt 420 gctgggcgca gccggccgcg gcacctcgcg agcacgccgg cctcggcagc gcgagcgcat 480 tagcgagatc tccgacgacg gacgtgggtc gagattcgat ttcggaggcc gcacgacgca 540 aaaaggtcat tcgagtttgt gtctgcggac tctgaacgtt cctcgtaaag cacttctga 599 <210> 31 <211> 500 <212> DNA <213> Cyanidioschyzon merolae (a red alga) <400> 31<s> 0002295 ccctcactgg catggcaaag ccgttctgtc tccgtgccgc atctgctcca ggggtgtaag 60 cgcgattgcg aatgctcaag gaaggttacg tgcacagtgg aatgcacgaa ataaccagta 120 catccgaaag gaagtacaag taatggaacc tgaaggtagg gtccagcagc atgatgggcg 180 ctttcccgaa tgtcaatacc gatctatcgc gcaatctggc ctccatggtc catagaagcg 240 ctttggcatc ggcgggagaa ccgggcgtcg ccccgcgctg cgctccatgg aacaatgctc 300 aaatcacgaa taaattgtac tttattaaat ctgtatgtac tatgatgtac aaaatagcat 360 tccaggaatc ccgagatcac acacgcgctc gagacgtgaa ctgtctcgtc actctcggct 420 acggcttcat cttcctcgta tttcttcgcc attagatacg agtgcggtaa gattctgggc 480 tgaagttttc aatattagtg 500 <210> 32 <211> 4700 <212> DNA <213> Artificial sequence <220> <223> pQE80 <400> 32 tagctgagct tggactcctg ttgatagatc cagtaatgac ctcagaactc catctggatt 60 tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaatcca agctagcttg 120 gcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc 180 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa 240 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa 300 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat 360 ccggaatttc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct 420 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac 480 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac 540 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg 600 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt 660 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag 720 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag 780 tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg gtgcccttaa 840 acgcctgggg taatgactct ctagcttgag gcatcaaata aaacgaaagg ctcagtcgaa 900 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa 960 tccgccctct agattacgtg cagtcgatga taagctgtca aacatgagaa ttgtgcctaa 1020 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 1080 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt 1140 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac 1200 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa 1260 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta 1320 tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc 1380 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag 1440 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat 1500 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga 1560 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg 1620 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg 1680 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc 1740 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt 1800 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct 1860 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag 1920 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc 1980 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt 2040 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc 2100 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc 2160 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcac cattcgatgg tgtcggaatt 2220 tcgggcagcg ttgggtcctg gccacgggtg cgcatgatct agagctgcct cgcgcgtttc 2280 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg 2340 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt 2400 cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg 2460 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat 2520 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc 2580 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 2640 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 2700 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 2760 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 2820 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 2880 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 2940 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 3000 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 3060 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 3120 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat 3180 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 3240 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 3300 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 3360 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 3420 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 3480 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 3540 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 3600 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 3660 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 3720 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 3780 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 3840 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 3900 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 3960 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 4020 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 4080 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 4140 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 4200 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 4260 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 4320 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 4380 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 4440 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta 4500 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcacctcg 4560 agaaatcata aaaaatttat ttgctttgtg agcggataac aattataata gattcaattg 4620 tgagcggata acaatttcac acagaattca ttaaagagga gaaattaact atgagaggat 4680 cgcatcacca tcaccatcac 4700 <210> 33 <211> 1986 <212> DNA <213> Cyanidioschyzon merolae (a red alga) <400> 33 gcgtgagtca gttcactgac aataagggaa acttgcgctt caaatgggag ccagccgagg 60 ccgtgttggc ggtggcattc gatgttcctg tgcccgggta tgatacatac aattgcatca 120 atctgcgctt gtgggacagt aagcctgcgc gtgagttcga tcttagctct ttcaacgttg 180 gcgactatta taagattctt gaaatgcggc agacgagtga gacgctctcc gccgtcctgt 240 acccgaacga tagcactgaa gcaggcaagg agctgcgcct caagcaacag tatttcttcg 300 tttcggcgac gctgcaggac atcattcgga ggttcctgaa gaaggaccga ccactcacgc 360 aacttgccga aaaagtgtgc attcagctga acgacacgca tccgacgatc gggattgttg 420 aaatgatgcg ccttctcctg gacgagtacg cattgggctg gacggatgcg tggaaaaccg 480 tcaaagcggt gttctcgtac acgaatcaca cggtgctgcc ggaggccttg gaaaagtggc 540 ctgtgccact catggaacgg ctcttgcctc gccacatgca gctcatcttc gaaatcaact 600 ttcgtcatct ccaggagtat gctcgtctaa gcaacaacga tggccatctg ttggagcgag 660 tgagcatcat cgaggagggt tttccaaaaa tggtgcgcat ggcccagctg gccgtcgttg 720 gctcacatac tgtcaatggt gtcgcggaaa tacactcgga actcgtgcga acccgtctct 780 tccctgattt taaccgcttc gagccgaaaa agtttgtgaa catcacaaac ggtgtgaccc 840 ctcgacgctg gatactggaa gcaaatcccg ccttgagtgc ggtgttttcc cgctggacgg 900 agagcgatga atggattttg gatttgaacc agatccgcca gttggaacag tacgccgaga 960 accctgacct gcaacgggaa tttttcgaag ccaaaaagga aaataagcgg cgtctcgctg 1020 aatacattcg agaaaagaat ggcgtccacg tggatgtgaa tgccctcttc gacatccagg 1080 tcaagcgaat tcacgagtac aagcgccagt tgttgaatat tctcggtgtg attgcgcgct 1140 acaatttgat caagtcgggc aagcgtgatc tcgtgccgcg ggtcttcatc ttcggaggca 1200 aagcagcggc tggttacgca caagccaagc gcatcattcg tctcattaat ggtgtagcag 1260 acgtggtcaa caatgatcca gatgttggcg acctcctgaa agttgtattt ctcgaaaact 1320 acagcgtgtc tcttgccgag atcatcattc cggcgagcga cattagcgag catatatcga 1380 ctgccggcat ggaggcgtca ggaacgagca acatgaagtt tgtgatgaac ggcggtctca 1440 tcatcggcac tatggacgga gcgaacattg aaatccgaga agaaatcggg ccggagaaca 1500 tctttattt cggtctgttg gctcaagaag ttgaccaggc gcgcaatgaa ctcaagtacc 1560 atggctggaa atgtaccgat gggcgcttcc agaacgcact gggtcagctc agtcgcggta 1620 tgtactgcgg tcaagacact tttcaggaaa ttgtacgagc cctcgatcca gccaacgact 1680 actacctgat cagtcgcgac tttacctcgt atatggaagc ccaggatcgc gtcgacgctg 1740 cctataggga ccagcgctcg tggctggcta agtgtattgt gagcacggct cgcatgggta 1800 agttcagttc tgatcgcagt atccatgagt acgcggagcg catttggcgc attgagccat 1860 gtgcctatac accgtccagc atcacataca aggaaccagt tgagggtgtc tcagagccca 1920 ccgcagacgg tacagcgccg aagacgacgc tccgaggaac gtaaacatca agtcccgagg 1980 gtgacg 1986 <210> 34 <211> 600 <212> DNA <213> Cyanidioschyzon merolae (a red alga) <400> 34 cgacgagaac gtataaggag tgcgcacggc gttttgttac aataccgata gatgagtttc 60 gaacatcgca ttcacaccat gagcgggggc gcacgctcca gagagtggag atggaaaagt 120 gccagcggag ccctgaggat gcaaaaaagt acggaccgct gacggaagag caaatggaaa 180 ggagggcgaa acttcgaggg ctacttgcat tagtaagtac aaccaacgat ggtaagaaac 240 gtatggagtt tgcgaaccga gactttaacg ctgccatcaa tatcaggaga tgtgcggtgc 300 tggagacgag acctccagag tgaacaagaa ggtacttttt tggacaacct tctaaggtcg 360 aactatatga gataaaattg gaagaagtag ttggtggccg gtccaaaaag acggggaggc 420 gtctgcacat cagttggaga cgttttgtcc aaggcgcgcc gatcgccact actgtacacg 480 gccggcgaga acgtggcgag aatacgcaag cgagctcgcc ggctgcgctc gctgcaccac 540 cgtcttttga ctcacaactt cgcgatatct ttgttctctg tgtttcttcg ttcgttgacc 600 <210> 35 <211> 717 <212> DNA <213> Cyanidioschyzon merolae (a red alga) <400> 35 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac 60 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac 120 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc 180 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag 240 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc 300 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg 360 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac 420 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac 480 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc 540 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaag 717 <210> 36 <211> 200 <212> DNA <213> Cyanidioschyzon merolae <400> 36 60. 60. 60. 60. 60. 60. 60. 60. 60. 60. 60. 60. 60 ggtaaaacac tcatgacgtt ttgtgttgaa ttgcaaccag cacgttatcg accagctctc 180. gacaaaaca tcgctttata cgggagaagt tgctgcggta ttgaccagag cgtacgaaaa cttgtgcagg caaaagtgt <210> 37 <211> 2757 <212> DNA <213> Cyanidioschyzon merolae <400> 37 60. gaactgaggg gcgaacgcag tcctccttgc attgttaagc aaaaaatatt ctctacaagc aatttgtgta caatctatat ggtacgctgc gtagagattt gcatgtcctg ctctacagtg cgaagaagca ttctatttta cgcagcggga gtaaagcaaa atcgagttga acgactatct 180 gacgcctacg caaagcgatt ccgcgtaccg tgtatctgca aagtagcact tcttaatggt 240 agagccgcaa ttgagaacgt tcggagtgtt ttcctggttc tgctggatgg atttttggta 300 aactatattt aatcttctag tgggaggatg attgcgcggg caaaaccata ttttccagga 360 tgagtcgtac tgaaagaagc acagtattca ctatgccggg cgaggtaggt gctagtttgg 420 gtttgacctc acgcgatgcg cacaacggct gcggaatacg cacaactcga tactgatcgt 480 cgagtcggca agactgcaac atgctcatgg ctcaggtcat ccgaagagaa acctcaccat 540 aaataccgtg ttcatacttc acatcgtaca taagaaaaat tacttttcag gccctctaca 600 ataataaaa ttggattcca cttatcgcgt ttatgagag atttaccgga cgcttccgtc 660 gcaaaaaaaa gaagagacat aagaaacttg acccactgtg tagaagga gcaaacaccg 720 caaagccctg cgaaaacctc atgcggtgat gtcgcgccct gctgcgatcg taacgaaggc 780 cattcgtcgc cacgcgcaac gcagggcaag agacggcttg cgtacggtgc agaacaccga 840 actggctgtg aagaggatct cttctttgtc agtactaaaa ggcgtcgctt tctgggaatg 900 gagcagcttg ccaaggatct ttttgagata ggtgcggtca agtttggcac gttcaaactc 960 aagtctggga tagcgtctcc attctatgtc gatctgcgcg ttgctgtctc gtatcctcga 1020 gttctgagaa gcattgcggc tctgtaccta gagtgcttgc aggacttttc tggcgctttc 1080 gacgttattt gcggggttcc atacactgcg ctcccttttg cgacagcaat ggctgtgcag 1140 ggagacttac ctatggttat gtgtcgaaag gaggtcaaag accatggac acggcgagtt 1200 gttgaaggtg ctttcacaca aggctctcgt tgcttaataa tcgaggacgt tgttacaagt 1260 1320 gcgatcgtct tgcttgatag ggagcaaggc gggtgtgagg cgcttctcgg ccagggaata 1380 gggctccgat ctgttttccg catcagcgac ctcgttggga cgctgggaa tttgaacttg 1440 cttgagccgg ggaaggtgga tatgattcta gatttattc gcacggcccg cgcagcatca 1500 gatgttaagc gccccgaaac aaacgttcca tccgcattgc cagtcgccgc atcaagttct 1560 tccccggtgc gagatcaact ttactcaatc gcagaaaaaa agagaagcat attatgcgtt 1620 gcggcagacg tacaatcgac taggagctt cttggaattg cagatgcagt aggcccccac 1680 atctgtgttc tgaagttgca cgccgatatt atccaggact ggaggacgga cacacaaga 1740 cgcttcgag agctcgcgga caaacagat ttcatgcttt ttgaggatcg aaaattcgct 1800 gacataggaa acacggttgt tgcacagttc agcgcaggcg tacatcggat ctcttcctgg 1860 gcggacattg tcaatgcaca cgccatcccg ggccctggac tgattgaggg cctgcgccac 1920 gcttgcgcaa tttccggcag aatgattggc ctcctgttag tcgcacaaat gtccagcaaa 1980 gggaacttga ttgacgagag atacactgct acatgcctgc ggatggcccg cgaagcgttt 2040 ccgttctgta tcggcttcat tgcccaggag cgtctggacc cgacagggaa gctctttgtg 2100 atggcgcctg gtgtgcaact ggattcaaag ggcgatcaat taggccaaca gtacaattca 2160 ccacagtatc tgctgaagcg caagggtgtc gacttcctta tcgttggccg cggtatctac 2220 gggagtgaag agccagcaaa ggcggcccag atgtacaaag agttgtcgtg gagcgtcctt 2280 cacggctaat tcctaatggg cagaagcaag cgtcggctcc cacaaagttc agtagaacgg cgggggtcag ttgccacgaa cgctttacgt gcggcatcaa gatgatattt cattgcgaat ggagttcgtc gacgtcagtt gcatccgcag aatgcttctt tgtttcggat atgtccgcgg 2460 cctgtgcccg caacgaatgt tcctagaagt acatttcacg gcgccttgaa catgtagtta agataagcgc tcgaaatcgt ccctgattcg gcagtgactg tccctagagc tggatcctat 2580. ggtttcttga ccctgtggct acaatatgcg ctcgacaccg tcgaagatca gggaatcgct ctagtgtcgg cctgatttcc accaagtgca ctctgcactt tttgaaattt ttggaggacc cattaaaaaa father ggattcctaa gaatagagat acagtcagct gctaggg <210> 38 <211> 1853 <212> DNA <213> Cyanidioschyzon merolae <400> 38 gaaaccgctc agcgaccag cgactcgagc tcgccacctc gtcgcgtggt ctcgtcgatg acctccttca tcgctaccaa gtagttcacg cgacgcaact tgcgcagatc ttttctcaga agaaacctca ggtcactcac atcggggtac tttcggccac gcgtgttcgc aacttcaaca 180 cagcgctgca cgacagcggt gaggtacgtc cgcaacaagt cctcgacaag ctcagccgta 240 tcacgccgcg gctgccgcgc atccccgaaa ccgtacagca tctggcgaat ttcattctgg 300 aataagcgct tcgaagcgtc ttcttcgggg tcgttcgcga aaaaagcatc ccagtcgttt 360 ccatcgctgg actgcatcgc actcgccgac tcggttccta gcacgaggcc gctgcctaga 420 caacgcgacg tacgcagata ctgtaagcaa taatagtgct gatgctgcca ctaacgaacc 480 tcgagcgcgt gtcagggcgc atacacgtcg ttaacgtacc gcggtggcag gcacggcgag 540 cttggcgtat gcgcgatacg taaatacatg ctgcgttgga tagcgactct gctgccgcat 600 ggcgacgctt tcatcagatg cggcgctgct gggaagaagg ccgtatccgg tgcatctgga 660 tggtagtttt gtataataat aggcttaaat gtcgttgcgt tcaaaggatg tgcaatcttt 720 ctggaagaaa tggaatatat gggacaaaaa agtacggaag gaaaagcatc tttaggtcca 780 tatatgaata gattggacac tgcatttcct gggtccaata acgccatcga acaaaatgga 840 tatatcggac aaaacatgta cggaacgtga agcaccgttc cctacgtcat caacactaga 900 cctcgtgttc tctcgccgat gctccggagc tatatgtacg ggatcgggcg tgcgcctgcg 960 cgttcctctt ctggacaggg cagcatctcg gtagcaacta ctgcagtgac tcacgcgtga 1020 tggtgtttgt tgcacctgtt ctacgacacg tgagcaagca ccgggcaagg agcaagcgac 1080 tggtgcgcgc aagcgtcagc akgacggaat ggtacgtgcc ggcgacactg gcccccaggg 1140 tgtcggtgaa taacacccag aatcttcgca actggagcgc gccagtgcga accgttcgcg 1200 tccctgtggc tacggtaggc gttggttccg ttgaacagca ccaaaagcac ctcggacacc 1260 catagacgtt ttggcttcgg cgcgagaccc gtctgccgga ctgggctacc ttgatgttgg 1320 tgtcagcgct tcctgttgtt gagctacgcg gtggcgtgcc ggtagggctc tggctcggtt 1380 tgtcgccggc cgagactttt ctcttctgcg ttatcggcaa catgcttcca ataccgtttc 1440 tagtttttgg tcttgcac gagcggatgc gtcgcctcgc gcgccccctg ttggactctg 1500 tggcgcggcg cctgccgagt catgcgaata ccccgtcgag tcaggcgctg gccctggcct 1560 tgtttgttgg cgtaccattg ccgggcactg gagcgtggtc tggcgctatt gccgcgttcc 1620 ttctgcaaat ggacatctgg cttgcgctgg tctccatcgc ggcgggcgtc gcgatcgctg 1680 gctgcatcat gattgcactc gttctaatgg gacgcatcgg tggactcatt gtcgcgtttg 1740 cgcttttagg agttggtgct agtgcgctgt ggcgcatgct gaaaccaccg tcgtcggcgg 1800 aaaactcgtg acatcaacgc cgccgcatct gagtgggtaa ggtcagcaag ctg 1853 <210> 39 <211> 200 <212> DNA <213> Cyanidioschyzon merolae (a red alga) <400> 39 cttatagctt acgtggcgga ttcgcagcga ttcgcgagat ttcccgaatc gccgatctcg 60 cgcgagatcc tcggcgaaga ttcgcggcgc atcatcgaac ggatcgcaga tgctgaaata 120 ccgcgcgcag cgaaattttt ccaacactag atttccattt gtgttctagc cgtggaaacc 180 tgtgagagaa ccagggattc 200 <210> 40 <211> 1209 <212> DNA <213> Cyanidioschyzon merolae (a red alga) <400> 40 atggtgttta cgtgtgctgc tttcgtagct ccagttggtg gttttcgcgg cacggcggtg 60 cgcgctacga gccgcgaggc cgtcggaccc cggctgcagg ctggtgacca gcctggtgcc 120 ttcactgcac gtacgcgaag cttaggtgtg ccgctgacgc ggagccggca acgtgctgcg 180 cactcgaatc ttgtgatgaa ggtgcgggta gcggtttccg ggttcggtcg catcggacgc 240 aactttgtgc gctgcctgca agcgaccccg aatgcgaacc tggagctggt ggggatcaac 300 gatacggctg gcatcaaaac tgccgctcat ctgctgaagt acgactctat tctgggaatt 360 gcaccgtttg acgtgaaggt tagcggggag agtaccatgc tcattgacgg gaagccggtt 420 accgttgtca gcaaccgcga tccgacccag ctgccttgga gggacctcaa cgtcgacatc 480 gtcatcgagg caacgggtgt ctttatctca agggacgggg caggcaagca catcgaagcg 540 ggtgccaaaa aggtggtcat cacagcgcca gcgaaaggcg aaggcgtccc gacctttgtc 600 atgggtctga acaacaccca gtataaccac gccaccgacc atgtggtgag caacgcgtcg 660 tgcaccacca acggcatggc accgttcgtg aaggtgctcg acgaggaatt cggcatcgtt 720 tccggcatga tgaccacgac gcattcgtat accggggacc agcgtctgct ggatgcatcg 780 catcgcgacc tgcgtcgcgc ccgtgcggcc gcgttgaaca tcgtgccgac ctcgacgggt 840 gccgcgcagg cggttgcgct ggtgtatcca ccagtgaagg gcaagctcac cggcattgcc 900 ctgcgagtgc cgaccccgaa cgtttctatc gtcgactttg tctgcacggt gaagaaacca 960 acgttcaagg aggaagtgaa tgcagctttc gtacgagcag cggaagggcc aatgaagggt 1020 atcctggcgg tgagcgatga gccgctcgtc tcatcggatt atcggatgaa cgtcaactca 1080 agcattgtgg atgcagcgct gacgacggtg atgggtgata cgctcgtcaa agtggtagcc 1140 tggtacgaca acgagtatgg ctacagtcaa cgggttgtgg acttggccaa ctacatagcg 1200 cagcatttc 1209 <210> 41 <211> 1572 <212> DNA <213> Artificial sequence <220> <223> codon optimized GP <400> 41 atggttccac aagcactgtt gctcgtgcct attctaggct tttccctgtg tttcgggaag 60 tttccgattt acaccattcc tgacacgttg ggcccatgga gtccgatcga catccatcac 120 ctgagctgcc caaacaactt ggtggttgag gatgaaggtt gcacgaatct ttccggtttc 180 tcgtatatgg aactgaaggt tggttacact agtgcgatca aggtgaacgg tttacatgc 240 aggaggtcg ttaccgaagc ggagacatat accaactttg taggctacgt gaccactacg 300 ttcaaacgga agcactttcg cccgactcct gatgcctgca gagctgctta caactggaag 360 atggcaggag atccgcgtta tgaggatcg ctccactccc cgtatccaga ttaccattgg 420 ctgcgaacag tgaagcacgaaggagagc ctggtcatca tatcgccctc agttgctgac 480 ctcgacccct atgacaatag cctgcattcg cgggtctttc ctagtggcaa atgcagcggg 540 ataacccgct cttcggtgta ctgctccact aaccacgact acacggtgtg gatgcctgag 600 atcttgcgac tcggcacgtc atgcgacatt ttcaccaaca gtcgaggcaa acgcgtgagc 660 aaagggaga cgacgtgtgg cttcatcgac gagaggggcc tctacaagtc actcaaaggg 720 gcctgcaagc tcaagctgtg tggcgttctt ggcctacgct tgatggatgg aacctgggta 780 tcaatgcaga catccaatga gacgaagtgg tgtccaccaa atcagctcgt aaatctccac 840 gacttacgct ccgatgaact ggaacatctg gtgattgaag agctcgtcaa gaaacgcgaa 900 gagtgcttag atgcgctcga gtccatcatt acgacgaaaa gcgtcagctt ccgccggctt 960 tcacacctgc gaaagctggt tcccggcttt ggtaaggcct acacgatctt caacaagacc 1020 ttgatggagg cagaagccca ctacaaatcg gtgcgtacct ggaacgagat cataccctct 1080 aaagggtgtc ttcgggtcgg aggtcgttgt catccgcatg tcaatggggt gtttttcaac 1140 ggcattatcc ttggtcccga tggtcacgtc ctgattccgg agatgcagtc gagcctcttg 1200 cagcagcata tcgagctcct ggagtcgtct gtgattccgc ttatgcatcc actcgcggat 1260 ccctttaccg tgttcaagga cggtgacgaa actgaggact tcatcgaggt ccacttaccg 1320 gacgtccacg aacaggtatc gggagtggat ctggggctgc ctaactgggg aaagtatgtg 1380 ctgcttagcg caggaacgct aatcgcgctc gttttgatca ttttcctaat gacctgttgc 1440 cgcaaggttg accggccgga aagtacgcaa cgctctctgc gtggtacagg ccgtaacgtc 1500 tcggttacgt cacaatcggg caaattcatc aacagctggg agtcgtacaa atccggtggc 1560 gaaacaggcc tt 1572 <210> 42 <211> 717 <212> DNA <213> Aequorea victoria <400> 42 atgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc 60 gacgtaaacg gccacaagtt cagcgtgcgt ggcgagggcg agggcgatgc caccaacggc 120 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc 180 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcgt 240 cacgacttct tcaagtccgc catgcccgaa ggctacgtc aggagcgcac catctcgttc 300 aaggacgacg gcacatacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg 360 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag 420 ctggagtaca actttaacag ccacaacgtc tatatcacag ccgacaagca gaagaacggc 480 atcaaggcaa acttcaagat ccgccacaac gttgaggacg gcagcgtgca gctcgccgac 540 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac 600 ctgagcaccc agtccgttct gagcaaagac cccaacgaga agcgcgatca catggtcctg 660 ctggagttcg tgaccgccgc cgggatcact cacggcatgg acgagctgta caagtaa 717 <210> 43 <211> 12 <212> PRT <213> Artificial sequence <220> <223> Co1 peptide <400> 43 Ser Phe His Gln Leu Pro Ala Arg Ser Pro Leu Pro 1 5 10 <210> 44 <211> 2781 <212> DNA <213> Cyanidioschyzon merolae (a red alga) <400> 44 gaactgaggg gcgaacgcag tcctccttgc attgttaagc aaaaaatatt ctctacaagc 60 aatttgtgta caatctatat ggtacgctgc gtagagattt gcatgtcctg ctctacagtg 120 cgaagaagca ttctatttta cgcagcggga gtaaagcaaa atcgagttga acgactatct 180 gacgcctacg caaagcgatt ccgcgtaccg tgtatctgca aagtagcact tcttaatggt 240 agagccgcaa ttgagaacgt tcggagtgtt ttcctggttc tgctggatgg atttttggta 300 aactatattt aatcttctag tgggaggatg attgcgcggg caaaacata ttttccagga 360 tgagtcgtac tgaaagagc acagtattca ctatgccggg cgaggtaggt gctagtttgg 420 gtttgacctc acgcgatgcg caaacggct gcggaatacg caaactcga tactgatcgt 480 cgagtcggca agactgcaac atgctcatgg ctcaggtcat ccgaagaa acctcaccat 540 aaataccgtg ttcatacttc acatcgtaca taagaaaaat tacttttcag gccctctaca 600 ataaaaaa ttggattcca cttatcgcgt ttattgagag atttaccgga cgcttccgtc 660 gcaaaaaaaa gaagagacat aagaaacttg acccactgtg tagagaga gcaaacaccg 720 caaagccctg cgaaaacctc atgcggtgat gtcgcgccct gctgcgatcg taacgaaggc 780 cattcgtcgc cacgcgcaac gcagggcaag agacggcttg cgtacggtgc agaacaccga 840 actggctgtg aagaggatct cttctttgtc agtactaaaa ggcgtcgctt tctgggaatg 900 gagcagcttg ccaaggatct ttttgagata ggtgcggtca agtttggcac gttcaaactc 960 aagtctggga tagcgtctcc attctatgtc gatctgcgcg ttgctgtctc gtatcctcga 1020 gttctgagaa gcattgcggc tctgtaccta gagtgcttgc aggacttttc tggcgctttc 1080 gacgttattt gcggggttcc atacactgcg ctcccttttg cgacagcaat ggctgtgcag 1140 ggagacttac ctatggttat gtgtcgaaag gaggtcaaag accatggaac acggcgagtt 1200 gttgaaggtg ctttcacaca aggctctcgt tgcttaataa tcgaggacgt tgttacaagt 1260 ggttcaagta ttctggaagt tgtgggcgca ttgcgcgcag agggtctcaa agtggatcac 1320 gcgatcgtct tgcttgatag ggagcaaggc gggtgtgagg cgcttctcgg ccagggaata 1380 gggctccgat ctgttttccg catcagcgac cttgtcggaa cactccgcca ctcaggtcgg 1440 ttatcagttg aacaagtaac cgagttgttt gattatttcc actcaaccaa agtatcatcg 1500 agtcctctga gtaactctat tcaaagtgga tataaagtgc atcctctttc ctttgaacaa 1560 cgtctttcat tgattcgaaa caaagttggc cgtcggctat tggaaattat gttaaagaag 1620 cagtcgaacc ttgcagtggc agcggatgta acaaccagtg aagaattatt atccattgcc 1680 aatgaagtgg gtccacaaat atgcatttta aagacacata tggatattat tcaagattgg 1740 acggaaagcg tatctgaaaa actcgttcat ttagctaagt tgcatcactt tttgatattt 1800 gaagacagaa agtttgcaga tattggcaat actgtggaat tacagttgac tggaggtata 1860 tttcatattg cacagtgggc agacatagtg aatgctcata tcattgctgg tcctggaact 1920 attcaagcgc taagtcgatc tgctgaacat tgcggtattt tgttattagc acaaatgagt 1980 agcaaaggga acttggcagt acaagaatat acgcaaaagg cattagaatt tgctcaacaa 2040 tatgaagatg ctgtttttgg ttttatatca ttgggttgta ttggagatcc aaactttctt 2100 tattttactc ctggagtgaa gttagaagga ggaggtgact ctttaggtca acaatatacg 2160 gatcctaaga cagttattgc tattcaagga agcgatgtag ctattgtagg gaggggaatt 2220 attcagtctt ctaatcgacg tgaagcagca tcaacttatc gaaaagcctg ttgggatgct 2280 tatttacaac gacttgaaga atacgttgaa taattcctaa tgggcagaag caagcgtcgg 2340 ctcccacaaa gttcagtaga acggcggggg tcagttgcca cgaacgcttt acgtgcggca 2400 tcaagatgat atttcattgc gaatggagtt cgtcgacgtc agttgcatcc gcagaatgct 2460 tctttgtttc ggatatgtcc gcggcctgtg cccgcaacga atgttcctag aagtacattt 2520 cacggcgcct tgaacatgta gttaagataa gcgctcgaaa tcgtccctga ttcggcagtg 2580 actgtcccta gagctggatc ctatggtttc ttgaccctgt ggctacaata tgcgctcgac 2640 accgtcgaag atcagggaat cgctctagtg tcggcctgat ttccaccaag tgcactctgc 2700 actttttgaa atttttggag gacccattaa aaaaataaat gcaaggattc ctaagaatag 2760 agatacagtc agctgctagg g 2781 <210> 45 <211> 390 <212> DNA <213> Cyanidioschyzon merolae (a red alga) <400> 45 atgttccatg tgacgtaccc gttcacgcag agacaatgct ttctccgttc acgagaagcg 60 tgccttgcaa cgttgccagc tggtgctttt cgaaagcacc tgtggcgccc ttcgtgctgg 120 tcgttccgca cacgtcttcg taaagaggcg tcgctacgga aatccacagt tctcgctccg 180 cttactcgcc gtctgcagct gagtctcttc ggcctcccag agcggttcgt tcgcaagtcc 240 aagtcgccgg tctcggcaga gtccagtgtc gccactgagc tcacacgtga tcgggtcaaa 300 gatccgacgc tcgcgagta ctgggataca cttctggaa tcaatgcact ggaggcgga ctggaacaac tcaaaagcga tgaactcaga <210> 46 <211> 90 <212> DNA <213> Removal (Artificial sequence) <220> <223> HA tag <400> 46 atgtacccat acgatgttcc tgactatgcg ggctatccct atgacgtccc ggactatgca ggataccctt atgacgttcc attacgct 90 <210> 47 <211> 30 <212> DNA <213> Removal (Artificial sequence) <220> <223> d184(+25)R <400> 47 cgtcaccctc gggacttgat gtttacgttc <210> 48 <211> 43 <212> DNA <213> Removal (Artificial sequence) <220> <223> bT3'(+1)F <400> 48 43. 43. 43. 43. 43. 43. 43. 43. 43. 43 <210> 49 <211> 39 <212> DNA <213> Artificial sequence <220> <223> HS(-200)Fd184 <400> 49 gtcccgaggg tgacgctttat agcttacgtg gcggattcg 39 <210> 50 <211> twenty two <212> DNA <213> Artificial sequence <220> <223> HS(-1)R <400> 50 gaatccctgg ttctctcaca gg 22 <210> 51 <211> 35 <212> DNA <213> Artificial sequence <220> <223> J042(1)Fhs <400> 51 gagaaccagg gattcatggt gtttacgtgt gctgc 35 <210> 52 <211> 41 <212> DNA <213> Artificial sequence <220> <223> J042(1209)R-link3 <400> 52 ggcgcctgca ccggatccga aatgctgcgc tatgtagttg g 41 <210> 53 <211> 35 <212> DNA <213> Artificial sequence <220> <223> GP(1)F-linker3 <400> 53 tccggtgcag gcgccatggt tccacaagca ctgtt 35 <210> 54 <211> 32 <212> DNA <213> Artificial sequence <220> <223> GP(1572)R-linker2 <400> 54 tccaccgcctccaccaaggcctgtttcgccac 32 <210> 55 <211> 35 <212> DNA <213> Artificial sequence <220> <223> sfGFP(1)F-linker2 <400> 55 ggtggaggcg gtggaggcat gagcaagggc gagga 35 <210> 56 <211> 35 <212> DNA <213> Artificial sequence <220> <223> sfGFP(714)Rbt <400> 56 taaatagcta gtttacttgt acagctcgtc catgc 35 <210> 57 <211> 28 <212> DNA <213> Artificial sequence <220> <223> D184(1200)F <400> 57 cgccttctcc tggacgagta cgcattgg <210> 58 <211> 21 <212> DNA <213> Removal (Artificial sequence) <220> <223> D184(+1400)R <400> 58 21. ccggccct accggcacgc c <210> 59 <211> 23 <212> DNA <213> Removal (Artificial sequence) <220> <223> APCC(-1)R <400> 59 ggtcaacga cgaagaaca cag <210> 60 <211> 43 <212> DNA <213> Removal (Artificial sequence) <220> <223> bT3'(+1) <400> 60 43. 43. 43. 43. 43. 43. 43. 43. 43. 43 <210> 61 <211> 35 <212> DNA <213> Removal (Artificial sequence) <220> <223> SecA(1)Fapcc <400> 61 cttcgttcgt tgaccatgtt ccatgtgacg taccc 35 <210> 62 <211> 90 <212> DNA <213> Removal (Artificial sequence) <220> <223> SecA(390)R-linker <400> 62 atcgtatggg tacatcccgg tgaacagctc ctcgcccttg ctcataccac cacctccgcc acctctgagt tcatcgcttt tgagttgttc <210> 63 <211> 32 <212> DNA <213> Removal (Artificial sequence) <220> <223> AND(1)F <400> 63 atgtacccat acgatgttcc tgactatgcg gg <210> 64 <211> 32 <212> DNA <213> Removal (Artificial sequence) <220> <223> AND(90)R <400> 64 agcgtaatct ggaacgtcat aagggtatcc tg <210> 65 <211> 37 <212> DNA <213> Removal (Artificial sequence) <220> <223> GP(1)Fha <400> 65 gttccagatt acgctatggt tccacaagca ctgttgc <210> 66 <211> 69 <212> DNA <213> Artificial sequence <220> <223> Co1-GP(1680)Rbt <400> 66 taaatagcta gtttatggga gcggcgagcg cgccggcagc tggtggaagc taaggcctgt 60 ttcgccacc 69

Claims

1. A drug delivery composition comprising acid-resistant cells encapsulating a drug, The acid-resistant cell is a haploid cell of an alga belonging to the class Cyanidiophyceae ( Cyanidiophyceae ), and is a cell that undergoes cell rupture under conditions of pH 7 or higher. wherein the drug is locally present in the chloroplast and is a drug that acts in the intestine.

2. The pharmaceutical delivery composition according to claim 1, wherein, The drug is at least one selected from the group consisting of low molecular weight compounds, peptides, proteins, and nucleic acids.

3. The pharmaceutical delivery composition according to claim 1 or 2, wherein The drug is a drug having immunogenicity.

4. The pharmaceutical delivery composition according to claim 1 or 2, wherein, The acid-resistant cells are cells resistant to acidic conditions of pH 1 to 3.

5. A feed for pets or experimental animals, which contains the drug delivery composition according to any one of claims 1 to 4.

6. A pharmaceutical product, which contains the drug delivery composition according to any one of claims 1 to 4.

7. A method for producing acid-resistant cells, which includes the step of introducing a gene encoding a fusion protein into acid-resistant cells, wherein the fusion protein contains a peptide or protein as a drug and a peptide or protein that is locally present relative to the chloroplast, The acid-resistant cell is a haploid cell of an alga belonging to the class Cyanidiophyceae( Cyanidiophyceae ) and is a cell that undergoes cell lysis under conditions with a pH of 7 or higher. wherein the drug is a drug that acts in the intestine.

8. A drug carrier comprising haploid cells of an alga belonging to the class Cyanidiophyceae ). The cells of the haploid of the alga are cells that undergo cell rupture under conditions of pH 7 or higher and are capable of encapsulating a drug, wherein the drug is locally present in the chloroplast and is a drug that acts in the intestine.

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