CN112028972A - Signal peptide for improving secretion expression of recombinant protein of mammalian cell and application thereof - Google Patents
Signal peptide for improving secretion expression of recombinant protein of mammalian cell and application thereof Download PDFInfo
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Abstract
The invention provides a signal peptide for improving secretion expression of recombinant protein of mammalian cells and application thereof. The present invention first provides a polypeptide of the following (a) or (b): (a) polypeptide consisting of an amino acid sequence shown in SEQ ID No. 1; (b) the polypeptide which is formed by substituting, deleting and/or adding one or more amino acids in the amino acid sequence shown in SEQ ID No.1 and has the same function with the polypeptide formed by the amino acid sequence shown in SEQ ID No. 1. The polypeptide can be used as a signal peptide to secrete and express a target protein in mammalian cells, is favorable for improving the secretion and expression level of the target protein, screens a high-expression monoclonal cell strain, and greatly reduces the time and cost for screening the high-expression signal peptide of the secreted protein.
Description
Technical Field
The present invention relates to a signal peptide and applications thereof, and more particularly, to a signal peptide for enhancing secretory expression of recombinant proteins in mammalian cells and applications thereof.
Background
Recombinant protein drugs are bright pearl in biological drugs, and with the progress of drug production technology and the increase of drug demand, the types of recombinant protein drugs in China are continuously abundant.
Systems for producing recombinant proteins using mammalian cells have been established for a long time. Compared with lower eukaryotic cell expression systems or prokaryotic expression systems, mammalian cell expression systems can guide the correct folding of proteins, provide various post-translational processing modification functions such as complex N-type glycosylation and accurate O-type glycosylation, so that the expressed product is closest to natural higher biological Protein molecules in terms of molecular structure, physicochemical properties and biological functions, and becomes the best choice for expressing recombinant proteins in the field of biopharmaceuticals (Chen P, Hutter D, Liu P, et al, Protein Expr Purif, 2002; 24 (3); 481-488).
One prominent feature of mammalian cell expression systems is the low level of protein expression. In recent years, with the development of molecular biology and Cell biology, a series of studies on the improvement of protein expression level have been carried out, including Cell strain modification (Davy AM, et al, Cell Syst.2017Mar 22; 4(3): 262-. Many researchers have done a lot of work on vector engineering, including selecting appropriate promoters, enhancers, signal peptides, all of which can increase protein expression levels in mammalian cells to some extent. Different signal peptides direct different levels of secretory protein expression, and selection of an appropriate signal peptide can increase the expression level of the protein (Steve Barash, et al, Biochem Biophys Res Commun.2002Jun 21; 294(4): 835-42).
Signal peptides are short peptide chains that direct the transfer of newly synthesized proteins to the secretory pathway, are located at the N-terminus of secreted proteins, can be cleaved, and typically consist of a 15-30 amino acid leader sequence, including three regions: a positively charged N-terminus of 2-3 polar amino acids, termed the basic amino terminus; an intermediate hydrophobic sequence, mainly neutral amino acids, capable of forming an alpha helical structure, which is the main functional region of the signal peptide; a longer, negatively charged C-terminus is the signal sequence cleavage site, also called the processing region. When the signal peptide sequence is synthesized, it is recognized by a Signal Recognition Particle (SRP), protein synthesis is suspended or slowed, the signal recognition particle carries the ribosome to the endoplasmic reticulum, protein synthesis is restarted, the newly synthesized protein enters the endoplasmic reticulum cavity under the guidance of the signal peptide, and the signal peptide sequence is cleaved by the action of a signal peptidase. The signal peptide is sufficient to transport any additional polypeptide to the target membrane. Different proteins are expressed in mammalian cells, different signal peptides are needed, and the secretion efficiency of the same protein under the action of different signal peptides is far from the same even if the protein can be secreted and expressed. For example, Jia-Ye Wang researches on the influence of mutating Pro at position 22 of a human tissue plasminogen activator signal peptide to Ala/Gly on the secretion effect of influenza virus H3N2-HA1, and the results show that compared with the non-mutated human tissue plasminogen activator signal peptide, the mutation of Pro at position 22 to Ala or Gly can effectively improve the secretion efficiency and expression level of H3N2-HA1 protein, wherein the mutation is the best Ala (Jia-Ye Wang et al, apple Microbiol Biotechnol.2011Aug; 91(3): 731-40). For secretory protein, the expression level can be improved by times by selecting proper signal peptide, and the method has a very profound significance for industrial production and scientific research.
In the prior art, if a higher expression level is required to be obtained, a plurality of signal peptides need to be screened separately for each secreted protein, and a proper signal peptide is selected from the signal peptides, so that the defects of long screening time, high cost and the like exist.
Disclosure of Invention
It is an object of the present invention to provide a novel signal peptide.
It is another object of the present invention to provide related uses of the signal peptide.
In one aspect, the present invention provides a polypeptide of (a) or (b):
(a) polypeptide consisting of an amino acid sequence shown in SEQ ID No. 1;
(b) the polypeptide which is formed by substituting, deleting and/or adding one or more amino acids in the amino acid sequence shown in SEQ ID No.1 and has the same function with the polypeptide formed by the amino acid sequence shown in SEQ ID No. 1.
The polypeptide of the invention is a signal peptide which can effectively improve the secretory expression of target proteins (including antibodies) in mammalian cells. In some specific embodiments of the invention, 9 verification proteins are selected to verify the function of the polypeptide as a signal peptide, and experimental results show that the expression level of the selected verification proteins is greatly improved by using the polypeptide consisting of the amino acid sequence shown in SEQ ID No.1 as the signal peptide, except that the expression level of the Human PD-1 is improved by about 4 times, the expression level of other verification proteins is improved by 5 times or more, and the expression level of more than 50% of the verification proteins is improved by 10 times or more. The HumanIL-17RC and the Human Decorin protein which are hardly expressed by the self signal peptide can successfully obtain the target protein under the condition of using the signal peptide, and the signal peptide can be widely used for the high-efficiency secretory expression of the target protein in mammalian cells and can effectively solve the production problem of the protein which is difficult to express.
According to a specific embodiment of the present invention, the polypeptide consisting of the amino acid sequence shown in SEQ ID No.1 with one or more amino acid substitutions, deletions and/or additions and having the same function as the polypeptide shown in SEQ ID No.1 at least comprises a sequence of at least 9 consecutive amino acids, preferably at least 10 consecutive amino acids, more preferably at least 11 consecutive amino acids at the C-terminal of the amino acid sequence shown in SEQ ID No. 1.
According to a specific embodiment of the present invention, the polypeptide having the same function as the polypeptide consisting of the amino acid sequence shown in SEQ ID No.1 refers to the polypeptide having the same function of increasing the secretory expression level of the target protein in the mammalian cells (compared to the use of the self-signal peptide, the secretory expression level of the target protein in the mammalian cells) as the polypeptide consisting of the amino acid sequence shown in SEQ ID No.1, preferably, having the same function of increasing the secretory expression level of the target protein in the mammalian cells by at least 4 times.
In another aspect, the invention also provides polynucleotides encoding the polypeptides of the invention.
The polynucleotide encoding the polypeptide of the present invention may be designed and prepared according to a conventional method.
In another aspect, the present invention also provides an expression vector comprising the polynucleotide of the present invention.
According to a specific embodiment of the present invention, the promoter of the expression vector of the present invention may be EF-1. alpha. promoter, hCMV (human cytokine viral) promoter, SV40(Simian vacuolating virus40) late promoter, or the like. In some embodiments of the invention, the expression vector is pcdan3.1 (+).
In another aspect, the present invention also provides a mammalian cell comprising the polynucleotide of the present invention or the expression vector of the present invention. The mammalian cell is selected from one or more of Chinese Hamster Ovary (CHO), mouse plasmacytoma (A non-Ig cloning, non-light chain-synthesizing of NS-1, NSO), Baby Hamster Kidney (BHK), human embryo kidney293 (HEK 293), etc.
In another aspect, the invention also provides the application of the polypeptide as a signal peptide in secretory expression of a target protein in mammalian cells.
According to a specific embodiment of the invention, the protein of interest can be selected from one or more of exogenous proteins such as Human IL-17RC, Human CD28, Human PD-1, Human ICOS, Human Decorin, Human Siglec-15, Human IL-7, Human IL-17F, and Human CD 73.
In another aspect, the present invention provides a method for secretory expression of a target protein in a mammalian cell, the method comprising:
the target protein is secreted and expressed by mammalian cells containing the polynucleotide or the expression vector.
According to a specific embodiment of the present invention, the method for secretory expression of a protein of interest in a mammalian cell of the present invention comprises:
linking a polynucleotide encoding the polypeptide of the present invention and a polynucleotide encoding a protein of interest, which is positioned at the front end of the polynucleotide encoding the protein of interest, and cloning the linked polynucleotide into a mammalian cell expression vector;
and (3) transfecting the expression vector into a mammalian cell to express the target protein.
According to a specific embodiment of the present invention, the method for secretory expression of a target protein in a mammalian cell of the present invention further comprises culturing the transfected mammalian cell to secretory expression of the target protein, and isolating the target protein from the culture.
In some specific embodiments of the present invention, 9 tumor immune drug development key target proteins or cytokines are selected as verification proteins of the application effect of the signal peptide of the present invention, and the amino acid sequences of the verification proteins are shown as SEQ ID nos. 11 to 19 in table 1. The amino acid sequence of the self signal peptide of the verification protein is shown as SEQ ID No. 2-SEQ ID No.10 in the table 1. In the invention, a section of poly His tag is introduced into the C terminal of the verification protein for separating and purifying the protein, and the amino acid sequence is shown as SEQ ID No.20 in Table 1. The amino acid sequences comprising the signal peptide, the verification protein and the poly His tag of the present invention are shown in SEQ ID No.21 to SEQ ID No.29 of Table 1. The amino acid sequences of the self-signal peptide, the verification protein and the poly His tag are shown in SEQ ID No. 30-SEQ ID No.38 in Table 1. The 9 selected tumor immune drugs are used for developing key target proteins or cytokines as foreign proteins to be expressed in HEK293 cells. Compared with the signal peptide of the protein self, the signal peptide of the invention is greatly improved in the expression level of the foreign protein in the HEK293 cell, except that the expression level of the Human PD-1 is improved by about 4 times, the expression level of other 8 verification proteins is improved by 5 times or more on average, and the expression level of more than 50% verification proteins is improved by 10 times or more.
In conclusion, the invention provides a novel signal peptide, which is applied to the improvement of the secretory expression level of secretory protein, screening of high-expression monoclonal cell strains and reduction of the production cost; on the other hand, the method greatly reduces the time and cost for screening the high-expression signal peptide of the secreted protein, and has wide application prospect.
Drawings
FIG. 1A is a map of expression vector pcDNA3.1, and FIG. 1B is a map of expression vector pEE14.4.
FIG. 2 is a schematic diagram of recombinant plasmid construction.
FIG. 3 shows the comparison of the effect of the signal peptide of the present invention and that of the self signal peptide.
FIG. 4 shows the results of comparison of the expression effect of Human protein in CHO cells using the signal peptide of the present invention and self signal peptide.
Detailed Description
In order that the invention may be more clearly understood, it will now be further described with reference to the following examples and the accompanying drawings. The examples are for illustration only and do not limit the invention in any way. The experimental methods in the examples, in which specific conditions are not noted, are conventional methods and conventional conditions well known in the art, or conditions as recommended by the manufacturer.
Example 1
In this example, 9 proteins were selected as verification proteins for the effect of the signal peptide of the present invention, and the signal peptide of the present invention and the self signal peptide of the verification protein were used to compare the secretion expression level of the verification protein in HEK293 cells under the same conditions. The specific operation is as follows:
1. construction of recombinant expression vectors
First, a recombinant expression vector using a signal peptide of the present invention (SEQ ID No.1, see Table 1 for specific sequence) was constructed, named pHEK1.0: synthesizing polynucleotides with amino acid sequences shown as SEQ ID No. 21-SEQ ID No.29 (the specific sequences are shown in Table 1) with sequences shown as SEQ ID No. 39-SEQ ID No.47 (wherein the polynucleotide sequence for encoding the signal peptide is shown as nucleotide sequences from 1 st to 51 st in SEQ ID No. 39-SEQ ID No. 47), and adding restriction enzyme BamH I cutting sites and Kozak sequences at the 5' tail end; the 3' end is added with restriction enzyme XhoI cutting site. The synthesized product is subjected to double enzyme digestion by restriction enzymes BamH I and Xho I, then agarose gel electrophoresis is carried out, and the gel is cut to recover the target fragment. The target fragment was ligated to pcNDA3.1(+) (purchased from Invitrogen) which was also double-digested with restriction enzymes BamHI and XhoI (expression vector pcDNA3.1 map is shown in FIG. 1A), recombinant eukaryotic expression vector pHEK1.0-IL-17RC/pHEK1.0-CD28/pHEK1.0-PD-1/pHEK1.0-ICOS/pHEK1.0-Decorin/pHEK1.0-Siglec-15/pHEK1.0-IL-7/pHEK1.0-IL-17F/pHEK1.0-CD73 (recombinant plasmid construction is shown in FIG. 2), E.coli DH5 alpha was transformed, cultured at 37 ℃ for 16h, single colony was picked up for plasmid extraction, the extracted plasmid was identified by double-restriction enzyme BamHI and XhoI, and then subjected to gel electrophoresis, and cloned in limited sea organism, and (4) selecting the clone with correct sequencing for amplifying and extracting the plasmid, and storing the extracted plasmid at the temperature of minus 20 ℃ for later use after sterile filtration.
Next, a recombinant expression vector using the verification protein self signal peptide was constructed, designated pNSP: synthesizing polynucleotides with amino acid sequences shown as SEQ ID No. 30-SEQ ID No.38 (the specific sequences are shown in Table 1) by Shanghai bioengineering Co., Ltd, wherein the sequences are SEQ ID No. 48-SEQ ID No.56, and restriction enzyme BamH I restriction sites and Kozak sequences are added at the 5' ends; the 3' end is added with restriction enzyme XhoI cutting site. The synthesized product is subjected to double enzyme digestion by restriction enzymes BamH I and Xho I, then agarose gel electrophoresis is carried out, and the gel is cut to recover the target fragment. Respectively connecting the target fragment with pcNDA3.1(+) (purchased from Invitrogen company) which is also subjected to double enzyme digestion by restriction enzymes BamHI and XhoI, constructing a recombinant eukaryotic expression vector pNSP-IL-17RC/pNSP-CD28/pNSP-PD-1/pNSP-ICOS/pNSP-Decorin/pNSP-Siglec-15/pNSP-IL-7/pNSP-IL-17F/pNSP-CD73, respectively converting Escherichia coli E.coli DH5 alpha, culturing at 37 ℃ for 16h, selecting a single colony for plasmid extraction, carrying out double agarose gel electrophoresis identification on the extracted plasmid after double enzyme digestion by the restriction enzymes BamHI and XhoI, sending the positive clone to Shanghai bioengineering company for sequencing, selecting a correct clone for amplified plasmid sequencing, carrying out sterile filtration and extracting the plasmid after extraction, storing at-20 deg.C for use.
TABLE 1
2. HEK293 cell transfected by recombinant expression vector and target protein expressed
And (3) transfecting HEK293 cells with the constructed recombinant plasmids by using a PEI transfection reagent respectively to express the target protein. HEK293 cells were plated at 1X 10 on the day before transfection6passage/mL, culture at 37 ℃. On day of transfection, counts were counted and adjustedCell density of 2X 106mL, survival rate above 95%.
The amounts of the components in the transfection reagent were calculated from the cell density: the corresponding relation between the plasmid dose and the number of cells is 1 × 106cells corresponded to 1. mu.g of plasmid, and the corresponding relationship between PEI dose and plasmid dose was such that the mass of PEI was 3 times the mass of DNA.
Preparing a transfection reagent according to the calculated component usage amount: 50 mu g of recombinant plasmid with the concentration of 200 mu g/mL is added into CD 293TGE culture medium to be 0.5mL (solution A), 150 mu g of PEI transfection reagent with the concentration of 1mg/mL is added into CD 293TGE culture medium to be 0.5mL (solution B), after respectively mixing uniformly, solution B is slowly added into solution A, after mixing uniformly and standing for 10-15min at room temperature, the transfection mixed solution is slowly dripped into 10mL HEK293 cells. 5% CO2After culturing at 37 ℃ and 135rpm for 96 hours, the supernatant was collected.
3. Separation and purification of target protein
And (3) separating and purifying the target protein by using a His-tag affinity chromatography method, and comparing the effect of the signal peptide of the invention with the effect of the signal peptide of the verification protein. The cell culture supernatant was collected by centrifugation at 1000 Xg for 30min, and the supernatant was filtered and bound to GE HisTrap excel chromatography columns equilibrated in advance with PBS, followed by elution of the target protein with PBS containing 30/100/500mM imidazole in an elution volume of 5CV per concentration, and the protein eluate was collected and subjected to SDS-PAGE analysis.
4. The signal peptide of the invention is used for analyzing the secretion expression effect of the test protein
Under the same conditions, the signal peptide of the invention and the self signal peptide of the verification protein are respectively used for comparing and verifying the secretion expression level of the protein in HEK293 cells, and the SDS-PAGE result is shown in figure 3. The results show that:
1) the signal peptide can improve the secretory expression quantity of the Human IL-17RC protein by more than 10 times;
2) the signal peptide can improve the secretory expression quantity of the Human CD28 protein by more than 10 times;
3) the signal peptide can improve the secretory expression quantity of the Human PD-1 protein by about 4 times;
4) the signal peptide can improve the secretory expression quantity of the Human ICOS protein by more than 5 times;
5) the signal peptide can improve the secretion expression quantity of the Human Decorin protein by more than 5 times;
6) the signal peptide can improve the secretion expression quantity of the Human Siglec-15 protein by about 10 times;
7) the signal peptide can improve the secretory expression quantity of the Human IL-7 protein by about 10 times;
8) the signal peptide can improve the secretory expression quantity of the Human IL-17F protein by about 5 times;
9) the signal peptide can improve the secretory expression quantity of the Human CD73 protein by more than 10 times.
Experimental data show that compared with the signal peptide of the protein, the signal peptide disclosed by the invention has the advantages that the expression quantity of the verification protein in the HEK293 cell is greatly improved, except that the expression quantity of human PD-1 is improved by about 4 times, the expression level of other verification proteins is improved by 5 times or more on average, and the expression level of over 50% verification proteins is improved by 10 times or more.
The results show that the signal peptide is particularly suitable for the secretion expression of the foreign protein of the HEK293 cell, the expression level of the verification protein is improved by 4 times or more, and the signal peptide can be widely used for the high-efficiency secretion expression of the foreign protein in the HEK293 cell.
Example 2
In the embodiment, the Human CD73 protein is selected as a verification protein for the application effect of the signal peptide in CHO cells. Under the same condition, the signal peptide of the invention and the self signal peptide of the verification protein are respectively used for comparing and verifying the secretion expression level of the Human CD73 protein in CHO cells. The specific operation is as follows:
1. construction of recombinant expression vectors
First, a recombinant expression vector using the signal peptide of the present invention (SEQ ID No.1, see Table 1 for a specific sequence) was constructed: the polynucleotide with the sequence of SEQ ID No.29 (the specific sequence is shown in Table 1) is synthesized by Shanghai bioengineering company Limited and has the sequence of SEQ ID No.47, and restriction enzyme Hind II I enzyme cutting site and Kozak sequence are added at the 5' end; the 3' end is added with a restriction enzyme EcoRI cutting site. The synthesized product is subjected to double digestion by restriction enzymes Hind I and EcoR I, then agarose gel electrophoresis is carried out, and the gel is cut to recover the target fragment. Respectively connecting the target fragment with pEE14.4 (purchased from Lonza biologics company) which is also subjected to double enzyme digestion by restriction enzymes HindII and EcoRI (the map of the expression vector pEE14.4 is shown in figure 1B), constructing a recombinant eukaryotic expression vector pCHO-CD73 (the recombinant plasmid is shown in figure 2), transforming Escherichia coli E.coli DH5 alpha, culturing at 37 ℃ for 16h, selecting a single colony for plasmid extraction, carrying out agarose gel electrophoresis identification after double enzyme digestion of the extracted plasmid by the restriction enzymes HindII and EcoRI, sending a positive clone to a marine bioengineering limited company for sequencing, selecting a clone with correct sequencing for amplification extraction of the plasmid, and preserving the extracted plasmid at-20 ℃ for later use after aseptic filtration.
Next, a recombinant expression vector using a signal peptide of the verification protein itself was constructed: the polynucleotide with the sequence of SEQ ID No.38 (the specific sequence is shown in Table 1) is synthesized by Shanghai bioengineering company Limited, the sequence is SEQ ID No.56, and restriction enzyme Hind II I enzyme cutting site and Kozak sequence are added at the 5' end; the 3' end is added with a restriction enzyme EcoRI cutting site. The synthesized product is subjected to double digestion by restriction enzymes Hind I and EcoR I, then agarose gel electrophoresis is carried out, and the gel is cut to recover the target fragment. Respectively connecting the target fragment with pEE14.4 (purchased from Lonza biologics company) which is also subjected to double enzyme digestion by restriction enzymes HindII and EcoRI, constructing a recombinant eukaryotic expression vector pNSP-CHO-CD73, transforming Escherichia coli E.coli DH5 alpha, culturing at 37 ℃ for 16h, selecting a single colony for plasmid extraction, carrying out double enzyme digestion on the extracted plasmid by the restriction enzymes HindII and EcoRI, carrying out agarose gel electrophoresis identification, sending the positive clone to Shanghai bioengineering Co., Ltd for sequencing, selecting the clone with correct sequencing for amplification extraction of the plasmid, and carrying out sterile filtration on the extracted plasmid and storing at-20 ℃ for later use.
2. Recombinant expression vector transfection CHO cell and expression of target protein
Expifeacmine was usedTMCHThe constructed recombinant plasmids pCHO-CD73 and pNSP-CHO-CD73 were transfected into CHO cells by the O Transfection Kit (available from Thermofeisher Co.) to express the target protein. CHO cells were plated at 1X 10 the day before transfection6passage/mL, culture at 37 ℃. Cells were counted on the day of transfection and adjusted to a cell density of 2X 106The activity rate is more than 95 percent.
The amounts of the components in the transfection reagent were calculated from the cell density: the corresponding relation between the plasmid dose and the number of cells is 1 × 106cells correspond to 1. mu.g plasmid, ExpifeacmineTMThe corresponding relation between the CHO Transfection Kit Transfection reagent and the plasmid dosage is that the mass of the Transfection reagent is 3 times of the DNA mass.
Preparing a transfection reagent according to the calculated component usage amount: 50. mu.g of the recombinant plasmid at a concentration of 200. mu.g/mL was added to OptiPROTMMedium (purchased from Thermofeisher Co.) to 0.5mL (solution A), 150. mu.g Expifactamine at a concentration of 1mg/mLTMAdding OptiPRO into CHO Transfection Kit Transfection reagentTMMedium Medium (purchased from Thermofeisher Co.) to 0.5mL (solution B), mixing, slowly adding solution B into solution A, mixing, standing at room temperature for 10min, and slowly dripping the transfection mixture into 10mL CHO cells. 5% CO2After culturing at 37 ℃ and 135rpm for 96 hours, the supernatant was collected.
3. Separation and purification of target protein
And (3) separating and purifying the target protein by using a His-tag affinity chromatography method, and comparing the effect of the signal peptide of the invention with the effect of the signal peptide of the verification protein. The cell culture supernatant was collected by centrifugation at 1000 Xg for 30min, and the supernatant was filtered and bound to GE HisTrap excel chromatography columns equilibrated in advance with PBS, followed by elution of the target protein with PBS containing 30/100/500mM imidazole in an elution volume of 5CV per concentration, and the protein eluate was collected and subjected to SDS-PAGE analysis.
4. The signal peptide of the invention is used for analyzing the secretion expression effect of the test protein
Under the same conditions, the secretion expression level of the protein HumanCD73 in CHO cells is verified by comparing the signal peptide of the invention and the self-signal peptide of the verification protein respectively, and the SDS-PAGE result is shown in FIG. 4. The results show that: the signal peptide can improve the secretion expression quantity of the Human CD73 protein in CHO cells by more than 8 times.
The results show that the signal peptide is not only particularly suitable for the secretion expression of the foreign protein of the HEK293 cell, but also can obviously improve the secretion expression level of the foreign protein in the CHO cell, and the signal peptide can be widely used for the high-efficiency secretion expression of the foreign protein in the mammalian cell.
SEQUENCE LISTING
<110> Beijing Baipusais Biotechnology Ltd
<120> Signal peptide for improving secretion expression of recombinant protein of mammalian cell and application thereof
<130> GAI20CN4660
<160> 56
<170> PatentIn version 3.5
<210> 1
<211> 17
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 1
Met Ser Ala Leu Leu Ile Leu Ala Leu Val Gly Ala Ala Val Val Trp
1 5 10 15
Ala
<210> 2
<211> 20
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 2
Met Pro Val Pro Trp Phe Leu Leu Ser Leu Ala Leu Gly Arg Ser Pro
1 5 10 15
Val Val Leu Ser
20
<210> 3
<211> 18
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 3
Met Leu Arg Leu Leu Leu Ala Leu Asn Leu Phe Pro Ser Ile Gln Val
1 5 10 15
Thr Gly
<210> 4
<211> 24
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 4
Met Gln Ile Pro Gln Ala Pro Trp Pro Val Val Trp Ala Val Leu Gln
1 5 10 15
Leu Gly Trp Arg Pro Gly Trp Phe
20
<210> 5
<211> 20
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 5
Met Lys Ser Gly Leu Trp Tyr Phe Phe Leu Phe Cys Leu Arg Ile Lys
1 5 10 15
Val Leu Thr Gly
20
<210> 6
<211> 16
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 6
Met Lys Ala Thr Ile Ile Leu Leu Leu Leu Ala Gln Val Ser Trp Ala
1 5 10 15
<210> 7
<211> 19
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 7
Met Glu Lys Ser Ile Trp Leu Leu Ala Cys Leu Ala Trp Val Leu Pro
1 5 10 15
Thr Gly Ser
<210> 8
<211> 25
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 8
Met Phe His Val Ser Phe Arg Tyr Ile Phe Gly Leu Pro Pro Leu Ile
1 5 10 15
Leu Val Leu Leu Pro Val Ala Ser Ser
20 25
<210> 9
<211> 30
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 9
Met Thr Val Lys Thr Leu His Gly Pro Ala Met Val Lys Tyr Leu Leu
1 5 10 15
Leu Ser Ile Leu Gly Leu Ala Phe Leu Ser Glu Ala Ala Ala
20 25 30
<210> 10
<211> 26
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 10
Met Cys Pro Arg Ala Ala Arg Ala Pro Ala Thr Leu Leu Leu Ala Leu
1 5 10 15
Gly Ala Val Leu Trp Pro Ala Ala Gly Ala
20 25
<210> 11
<211> 445
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 11
Leu Glu Arg Leu Val Gly Pro Gln Asp Ala Thr His Cys Ser Pro Gly
1 5 10 15
Leu Ser Cys Arg Leu Trp Asp Ser Asp Ile Leu Cys Leu Pro Gly Asp
20 25 30
Ile Val Pro Ala Pro Gly Pro Val Leu Ala Pro Thr His Leu Gln Thr
35 40 45
Glu Leu Val Leu Arg Cys Gln Lys Glu Thr Asp Cys Asp Leu Cys Leu
50 55 60
Arg Val Ala Val His Leu Ala Val His Gly His Trp Glu Glu Pro Glu
65 70 75 80
Asp Glu Glu Lys Phe Gly Gly Ala Ala Asp Ser Gly Val Glu Glu Pro
85 90 95
Arg Asn Ala Ser Leu Gln Ala Gln Val Val Leu Ser Phe Gln Ala Tyr
100 105 110
Pro Thr Ala Arg Cys Val Leu Leu Glu Val Gln Val Pro Ala Ala Leu
115 120 125
Val Gln Phe Gly Gln Ser Val Gly Ser Val Val Tyr Asp Cys Phe Glu
130 135 140
Ala Ala Leu Gly Ser Glu Val Arg Ile Trp Ser Tyr Thr Gln Pro Arg
145 150 155 160
Tyr Glu Lys Glu Leu Asn His Thr Gln Gln Leu Pro Asp Cys Arg Gly
165 170 175
Leu Glu Val Trp Asn Ser Ile Pro Ser Cys Trp Ala Leu Pro Trp Leu
180 185 190
Asn Val Ser Ala Asp Gly Asp Asn Val His Leu Val Leu Asn Val Ser
195 200 205
Glu Glu Gln His Phe Gly Leu Ser Leu Tyr Trp Asn Gln Val Gln Gly
210 215 220
Pro Pro Lys Pro Arg Trp His Lys Asn Leu Thr Gly Pro Gln Ile Ile
225 230 235 240
Thr Leu Asn His Thr Asp Leu Val Pro Cys Leu Cys Ile Gln Val Trp
245 250 255
Pro Leu Glu Pro Asp Ser Val Arg Thr Asn Ile Cys Pro Phe Arg Glu
260 265 270
Asp Pro Arg Ala His Gln Asn Leu Trp Gln Ala Ala Arg Leu Gln Leu
275 280 285
Leu Thr Leu Gln Ser Trp Leu Leu Asp Ala Pro Cys Ser Leu Pro Ala
290 295 300
Glu Ala Ala Leu Cys Trp Arg Ala Pro Gly Gly Asp Pro Cys Gln Pro
305 310 315 320
Leu Val Pro Pro Leu Ser Trp Glu Asn Val Thr Val Asp Lys Val Leu
325 330 335
Glu Phe Pro Leu Leu Lys Gly His Pro Asn Leu Cys Val Gln Val Asn
340 345 350
Ser Ser Glu Lys Leu Gln Leu Gln Glu Cys Leu Trp Ala Asp Ser Leu
355 360 365
Gly Pro Leu Lys Asp Asp Val Leu Leu Leu Glu Thr Arg Gly Pro Gln
370 375 380
Asp Asn Arg Ser Leu Cys Ala Leu Glu Pro Ser Gly Cys Thr Ser Leu
385 390 395 400
Pro Ser Lys Ala Ser Thr Arg Ala Ala Arg Leu Gly Glu Tyr Leu Leu
405 410 415
Gln Asp Leu Gln Ser Gly Gln Cys Leu Gln Leu Trp Asp Asp Asp Leu
420 425 430
Gly Ala Leu Trp Ala Cys Pro Met Asp Lys Tyr Ile His
435 440 445
<210> 12
<211> 134
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 12
Asn Lys Ile Leu Val Lys Gln Ser Pro Met Leu Val Ala Tyr Asp Asn
1 5 10 15
Ala Val Asn Leu Ser Cys Lys Tyr Ser Tyr Asn Leu Phe Ser Arg Glu
20 25 30
Phe Arg Ala Ser Leu His Lys Gly Leu Asp Ser Ala Val Glu Val Cys
35 40 45
Val Val Tyr Gly Asn Tyr Ser Gln Gln Leu Gln Val Tyr Ser Lys Thr
50 55 60
Gly Phe Asn Cys Asp Gly Lys Leu Gly Asn Glu Ser Val Thr Phe Tyr
65 70 75 80
Leu Gln Asn Leu Tyr Val Asn Gln Thr Asp Ile Tyr Phe Cys Lys Ile
85 90 95
Glu Val Met Tyr Pro Pro Pro Tyr Leu Asp Asn Glu Lys Ser Asn Gly
100 105 110
Thr Ile Ile His Val Lys Gly Lys His Leu Cys Pro Ser Pro Leu Phe
115 120 125
Pro Gly Pro Ser Lys Pro
130
<210> 13
<211> 143
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 13
Leu Asp Ser Pro Asp Arg Pro Trp Asn Pro Pro Thr Phe Ser Pro Ala
1 5 10 15
Leu Leu Val Val Thr Glu Gly Asp Asn Ala Thr Phe Thr Cys Ser Phe
20 25 30
Ser Asn Thr Ser Glu Ser Phe Val Leu Asn Trp Tyr Arg Met Ser Pro
35 40 45
Ser Asn Gln Thr Asp Lys Leu Ala Ala Phe Pro Glu Asp Arg Ser Gln
50 55 60
Pro Gly Gln Asp Cys Arg Phe Arg Val Thr Gln Leu Pro Asn Gly Arg
65 70 75 80
Asp Phe His Met Ser Val Val Arg Ala Arg Arg Asn Asp Ser Gly Thr
85 90 95
Tyr Leu Cys Gly Ala Ile Ser Leu Ala Pro Lys Ala Gln Ile Lys Glu
100 105 110
Ser Leu Arg Ala Glu Leu Arg Val Thr Glu Arg Arg Ala Glu Val Pro
115 120 125
Thr Ala His Pro Ser Pro Ser Pro Arg Pro Ala Gly Gln Phe Gln
130 135 140
<210> 14
<211> 120
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 14
Glu Ile Asn Gly Ser Ala Asn Tyr Glu Met Phe Ile Phe His Asn Gly
1 5 10 15
Gly Val Gln Ile Leu Cys Lys Tyr Pro Asp Ile Val Gln Gln Phe Lys
20 25 30
Met Gln Leu Leu Lys Gly Gly Gln Ile Leu Cys Asp Leu Thr Lys Thr
35 40 45
Lys Gly Ser Gly Asn Thr Val Ser Ile Lys Ser Leu Lys Phe Cys His
50 55 60
Ser Gln Leu Ser Asn Asn Ser Val Ser Phe Phe Leu Tyr Asn Leu Asp
65 70 75 80
His Ser His Ala Asn Tyr Tyr Phe Cys Asn Leu Ser Ile Phe Asp Pro
85 90 95
Pro Pro Phe Lys Val Thr Leu Thr Gly Gly Tyr Leu His Ile Tyr Glu
100 105 110
Ser Gln Leu Cys Cys Gln Leu Lys
115 120
<210> 15
<211> 343
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 15
Gly Pro Phe Gln Gln Arg Gly Leu Phe Asp Phe Met Leu Glu Asp Glu
1 5 10 15
Ala Ser Gly Ile Gly Pro Glu Val Pro Asp Asp Arg Asp Phe Glu Pro
20 25 30
Ser Leu Gly Pro Val Cys Pro Phe Arg Cys Gln Cys His Leu Arg Val
35 40 45
Val Gln Cys Ser Asp Leu Gly Leu Asp Lys Val Pro Lys Asp Leu Pro
50 55 60
Pro Asp Thr Thr Leu Leu Asp Leu Gln Asn Asn Lys Ile Thr Glu Ile
65 70 75 80
Lys Asp Gly Asp Phe Lys Asn Leu Lys Asn Leu His Ala Leu Ile Leu
85 90 95
Val Asn Asn Lys Ile Ser Lys Val Ser Pro Gly Ala Phe Thr Pro Leu
100 105 110
Val Lys Leu Glu Arg Leu Tyr Leu Ser Lys Asn Gln Leu Lys Glu Leu
115 120 125
Pro Glu Lys Met Pro Lys Thr Leu Gln Glu Leu Arg Ala His Glu Asn
130 135 140
Glu Ile Thr Lys Val Arg Lys Val Thr Phe Asn Gly Leu Asn Gln Met
145 150 155 160
Ile Val Ile Glu Leu Gly Thr Asn Pro Leu Lys Ser Ser Gly Ile Glu
165 170 175
Asn Gly Ala Phe Gln Gly Met Lys Lys Leu Ser Tyr Ile Arg Ile Ala
180 185 190
Asp Thr Asn Ile Thr Ser Ile Pro Gln Gly Leu Pro Pro Ser Leu Thr
195 200 205
Glu Leu His Leu Asp Gly Asn Lys Ile Ser Arg Val Asp Ala Ala Ser
210 215 220
Leu Lys Gly Leu Asn Asn Leu Ala Lys Leu Gly Leu Ser Phe Asn Ser
225 230 235 240
Ile Ser Ala Val Asp Asn Gly Ser Leu Ala Asn Thr Pro His Leu Arg
245 250 255
Glu Leu His Leu Asp Asn Asn Lys Leu Thr Arg Val Pro Gly Gly Leu
260 265 270
Ala Glu His Lys Tyr Ile Gln Val Val Tyr Leu His Asn Asn Asn Ile
275 280 285
Ser Val Val Gly Ser Ser Asp Phe Cys Pro Pro Gly His Asn Thr Lys
290 295 300
Lys Ala Ser Tyr Ser Gly Val Ser Leu Phe Ser Asn Pro Val Gln Tyr
305 310 315 320
Trp Glu Ile Gln Pro Ser Thr Phe Arg Cys Val Tyr Val Arg Ser Ala
325 330 335
Ile Gln Leu Gly Asn Tyr Lys
340
<210> 16
<211> 242
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 16
Phe Val Arg Thr Lys Ile Asp Thr Thr Glu Asn Leu Leu Asn Thr Glu
1 5 10 15
Val His Ser Ser Pro Ala Gln Arg Trp Ser Met Gln Val Pro Pro Glu
20 25 30
Val Ser Ala Glu Ala Gly Asp Ala Ala Val Leu Pro Cys Thr Phe Thr
35 40 45
His Pro His Arg His Tyr Asp Gly Pro Leu Thr Ala Ile Trp Arg Ala
50 55 60
Gly Glu Pro Tyr Ala Gly Pro Gln Val Phe Arg Cys Ala Ala Ala Arg
65 70 75 80
Gly Ser Glu Leu Cys Gln Thr Ala Leu Ser His Gly Arg Phe Arg Leu
85 90 95
Leu Gly Asn Pro Arg Arg Asn Asp Leu Ser Leu Arg Val Glu Arg Leu
100 105 110
Ala Leu Ala Asp Asp Arg Arg Tyr Phe Cys Arg Val Glu Phe Ala Gly
115 120 125
Asp Val His Asp Arg Tyr Glu Ser Arg His Gly Val Arg Leu His Val
130 135 140
Thr Ala Ala Pro Arg Ile Val Asn Ile Ser Val Leu Pro Ser Pro Ala
145 150 155 160
His Ala Phe Arg Ala Leu Cys Thr Ala Glu Gly Glu Pro Pro Pro Ala
165 170 175
Leu Ala Trp Ser Gly Pro Ala Leu Gly Asn Ser Leu Ala Ala Val Arg
180 185 190
Ser Pro Arg Glu Gly His Gly His Leu Val Thr Ala Glu Leu Pro Ala
195 200 205
Leu Thr His Asp Gly Arg Tyr Thr Cys Thr Ala Ala Asn Ser Leu Arg
210 215 220
Ser Glu Ala Ser Val Tyr Leu Phe Arg Phe His Gly Ala Ser Gly Ala
225 230 235 240
Ser Thr
<210> 17
<211> 152
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 17
Asp Cys Asp Ile Glu Gly Lys Asp Gly Lys Gln Tyr Glu Ser Val Leu
1 5 10 15
Met Val Ser Ile Asp Gln Leu Leu Asp Ser Met Lys Glu Ile Gly Ser
20 25 30
Asn Cys Leu Asn Asn Glu Phe Asn Phe Phe Lys Arg His Ile Cys Asp
35 40 45
Ala Asn Lys Glu Gly Met Phe Leu Phe Arg Ala Ala Arg Lys Leu Arg
50 55 60
Gln Phe Leu Lys Met Asn Ser Thr Gly Asp Phe Asp Leu His Leu Leu
65 70 75 80
Lys Val Ser Glu Gly Thr Thr Ile Leu Leu Asn Cys Thr Gly Gln Val
85 90 95
Lys Gly Arg Lys Pro Ala Ala Leu Gly Glu Ala Gln Pro Thr Lys Ser
100 105 110
Leu Glu Glu Asn Lys Ser Leu Lys Glu Gln Lys Lys Leu Asn Asp Leu
115 120 125
Cys Phe Leu Lys Arg Leu Leu Gln Glu Ile Lys Thr Cys Trp Asn Lys
130 135 140
Ile Leu Met Gly Thr Lys Glu His
145 150
<210> 18
<211> 133
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 18
Arg Lys Ile Pro Lys Val Gly His Thr Phe Phe Gln Lys Pro Glu Ser
1 5 10 15
Cys Pro Pro Val Pro Gly Gly Ser Met Lys Leu Asp Ile Gly Ile Ile
20 25 30
Asn Glu Asn Gln Arg Val Ser Met Ser Arg Asn Ile Glu Ser Arg Ser
35 40 45
Thr Ser Pro Trp Asn Tyr Thr Val Thr Trp Asp Pro Asn Arg Tyr Pro
50 55 60
Ser Glu Val Val Gln Ala Gln Cys Arg Asn Leu Gly Cys Ile Asn Ala
65 70 75 80
Gln Gly Lys Glu Asp Ile Ser Met Asn Ser Val Pro Ile Gln Gln Glu
85 90 95
Thr Leu Val Val Arg Arg Lys His Gln Gly Cys Ser Val Ser Phe Gln
100 105 110
Leu Glu Lys Val Leu Val Thr Val Gly Cys Thr Cys Val Thr Pro Val
115 120 125
Ile His His Val Gln
130
<210> 19
<211> 522
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 19
Trp Glu Leu Thr Ile Leu His Thr Asn Asp Val His Ser Arg Leu Glu
1 5 10 15
Gln Thr Ser Glu Asp Ser Ser Lys Cys Val Asn Ala Ser Arg Cys Met
20 25 30
Gly Gly Val Ala Arg Leu Phe Thr Lys Val Gln Gln Ile Arg Arg Ala
35 40 45
Glu Pro Asn Val Leu Leu Leu Asp Ala Gly Asp Gln Tyr Gln Gly Thr
50 55 60
Ile Trp Phe Thr Val Tyr Lys Gly Ala Glu Val Ala His Phe Met Asn
65 70 75 80
Ala Leu Arg Tyr Asp Ala Met Ala Leu Gly Asn His Glu Phe Asp Asn
85 90 95
Gly Val Glu Gly Leu Ile Glu Pro Leu Leu Lys Glu Ala Lys Phe Pro
100 105 110
Ile Leu Ser Ala Asn Ile Lys Ala Lys Gly Pro Ala Ser Gln Ile Ser
115 120 125
Gly Leu Tyr Leu Pro Tyr Lys Val Leu Pro Val Gly Asp Glu Val Val
130 135 140
Gly Ile Val Gly Tyr Thr Ser Lys Glu Thr Pro Phe Leu Ser Asn Pro
145 150 155 160
Gly Thr Asn Leu Val Phe Glu Asp Glu Ile Thr Ala Leu Gln Pro Glu
165 170 175
Val Asp Lys Leu Lys Thr Leu Asn Val Asn Lys Ile Ile Ala Leu Gly
180 185 190
His Ser Gly Phe Glu Met Asp Lys Leu Ile Ala Gln Lys Val Arg Gly
195 200 205
Val Asp Val Val Val Gly Gly His Ser Asn Thr Phe Leu Tyr Thr Gly
210 215 220
Asn Pro Pro Ser Lys Glu Val Pro Ala Gly Lys Tyr Pro Phe Ile Val
225 230 235 240
Thr Ser Asp Asp Gly Arg Lys Val Pro Val Val Gln Ala Tyr Ala Phe
245 250 255
Gly Lys Tyr Leu Gly Tyr Leu Lys Ile Glu Phe Asp Glu Arg Gly Asn
260 265 270
Val Ile Ser Ser His Gly Asn Pro Ile Leu Leu Asn Ser Ser Ile Pro
275 280 285
Glu Asp Pro Ser Ile Lys Ala Asp Ile Asn Lys Trp Arg Ile Lys Leu
290 295 300
Asp Asn Tyr Ser Thr Gln Glu Leu Gly Lys Thr Ile Val Tyr Leu Asp
305 310 315 320
Gly Ser Ser Gln Ser Cys Arg Phe Arg Glu Cys Asn Met Gly Asn Leu
325 330 335
Ile Cys Asp Ala Met Ile Asn Asn Asn Leu Arg His Thr Asp Glu Met
340 345 350
Phe Trp Asn His Val Ser Met Cys Ile Leu Asn Gly Gly Gly Ile Arg
355 360 365
Ser Pro Ile Asp Glu Arg Asn Asn Gly Thr Ile Thr Trp Glu Asn Leu
370 375 380
Ala Ala Val Leu Pro Phe Gly Gly Thr Phe Asp Leu Val Gln Leu Lys
385 390 395 400
Gly Ser Thr Leu Lys Lys Ala Phe Glu His Ser Val His Arg Tyr Gly
405 410 415
Gln Ser Thr Gly Glu Phe Leu Gln Val Gly Gly Ile His Val Val Tyr
420 425 430
Asp Leu Ser Arg Lys Pro Gly Asp Arg Val Val Lys Leu Asp Val Leu
435 440 445
Cys Thr Lys Cys Arg Val Pro Ser Tyr Asp Pro Leu Lys Met Asp Glu
450 455 460
Val Tyr Lys Val Ile Leu Pro Asn Phe Leu Ala Asn Gly Gly Asp Gly
465 470 475 480
Phe Gln Met Ile Lys Asp Glu Leu Leu Arg His Asp Ser Gly Asp Gln
485 490 495
Asp Ile Asn Val Val Ser Thr Tyr Ile Ser Lys Met Lys Val Ile Tyr
500 505 510
Pro Ala Val Glu Gly Arg Ile Lys Phe Ser
515 520
<210> 20
<211> 8
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 20
Gly Ser His His His His His His
1 5
<210> 21
<211> 470
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 21
Met Ser Ala Leu Leu Ile Leu Ala Leu Val Gly Ala Ala Val Val Trp
1 5 10 15
Ala Leu Glu Arg Leu Val Gly Pro Gln Asp Ala Thr His Cys Ser Pro
20 25 30
Gly Leu Ser Cys Arg Leu Trp Asp Ser Asp Ile Leu Cys Leu Pro Gly
35 40 45
Asp Ile Val Pro Ala Pro Gly Pro Val Leu Ala Pro Thr His Leu Gln
50 55 60
Thr Glu Leu Val Leu Arg Cys Gln Lys Glu Thr Asp Cys Asp Leu Cys
65 70 75 80
Leu Arg Val Ala Val His Leu Ala Val His Gly His Trp Glu Glu Pro
85 90 95
Glu Asp Glu Glu Lys Phe Gly Gly Ala Ala Asp Ser Gly Val Glu Glu
100 105 110
Pro Arg Asn Ala Ser Leu Gln Ala Gln Val Val Leu Ser Phe Gln Ala
115 120 125
Tyr Pro Thr Ala Arg Cys Val Leu Leu Glu Val Gln Val Pro Ala Ala
130 135 140
Leu Val Gln Phe Gly Gln Ser Val Gly Ser Val Val Tyr Asp Cys Phe
145 150 155 160
Glu Ala Ala Leu Gly Ser Glu Val Arg Ile Trp Ser Tyr Thr Gln Pro
165 170 175
Arg Tyr Glu Lys Glu Leu Asn His Thr Gln Gln Leu Pro Asp Cys Arg
180 185 190
Gly Leu Glu Val Trp Asn Ser Ile Pro Ser Cys Trp Ala Leu Pro Trp
195 200 205
Leu Asn Val Ser Ala Asp Gly Asp Asn Val His Leu Val Leu Asn Val
210 215 220
Ser Glu Glu Gln His Phe Gly Leu Ser Leu Tyr Trp Asn Gln Val Gln
225 230 235 240
Gly Pro Pro Lys Pro Arg Trp His Lys Asn Leu Thr Gly Pro Gln Ile
245 250 255
Ile Thr Leu Asn His Thr Asp Leu Val Pro Cys Leu Cys Ile Gln Val
260 265 270
Trp Pro Leu Glu Pro Asp Ser Val Arg Thr Asn Ile Cys Pro Phe Arg
275 280 285
Glu Asp Pro Arg Ala His Gln Asn Leu Trp Gln Ala Ala Arg Leu Gln
290 295 300
Leu Leu Thr Leu Gln Ser Trp Leu Leu Asp Ala Pro Cys Ser Leu Pro
305 310 315 320
Ala Glu Ala Ala Leu Cys Trp Arg Ala Pro Gly Gly Asp Pro Cys Gln
325 330 335
Pro Leu Val Pro Pro Leu Ser Trp Glu Asn Val Thr Val Asp Lys Val
340 345 350
Leu Glu Phe Pro Leu Leu Lys Gly His Pro Asn Leu Cys Val Gln Val
355 360 365
Asn Ser Ser Glu Lys Leu Gln Leu Gln Glu Cys Leu Trp Ala Asp Ser
370 375 380
Leu Gly Pro Leu Lys Asp Asp Val Leu Leu Leu Glu Thr Arg Gly Pro
385 390 395 400
Gln Asp Asn Arg Ser Leu Cys Ala Leu Glu Pro Ser Gly Cys Thr Ser
405 410 415
Leu Pro Ser Lys Ala Ser Thr Arg Ala Ala Arg Leu Gly Glu Tyr Leu
420 425 430
Leu Gln Asp Leu Gln Ser Gly Gln Cys Leu Gln Leu Trp Asp Asp Asp
435 440 445
Leu Gly Ala Leu Trp Ala Cys Pro Met Asp Lys Tyr Ile His Gly Ser
450 455 460
His His His His His His
465 470
<210> 22
<211> 159
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 22
Met Ser Ala Leu Leu Ile Leu Ala Leu Val Gly Ala Ala Val Val Trp
1 5 10 15
Ala Asn Lys Ile Leu Val Lys Gln Ser Pro Met Leu Val Ala Tyr Asp
20 25 30
Asn Ala Val Asn Leu Ser Cys Lys Tyr Ser Tyr Asn Leu Phe Ser Arg
35 40 45
Glu Phe Arg Ala Ser Leu His Lys Gly Leu Asp Ser Ala Val Glu Val
50 55 60
Cys Val Val Tyr Gly Asn Tyr Ser Gln Gln Leu Gln Val Tyr Ser Lys
65 70 75 80
Thr Gly Phe Asn Cys Asp Gly Lys Leu Gly Asn Glu Ser Val Thr Phe
85 90 95
Tyr Leu Gln Asn Leu Tyr Val Asn Gln Thr Asp Ile Tyr Phe Cys Lys
100 105 110
Ile Glu Val Met Tyr Pro Pro Pro Tyr Leu Asp Asn Glu Lys Ser Asn
115 120 125
Gly Thr Ile Ile His Val Lys Gly Lys His Leu Cys Pro Ser Pro Leu
130 135 140
Phe Pro Gly Pro Ser Lys Pro Gly Ser His His His His His His
145 150 155
<210> 23
<211> 168
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 23
Met Ser Ala Leu Leu Ile Leu Ala Leu Val Gly Ala Ala Val Val Trp
1 5 10 15
Ala Leu Asp Ser Pro Asp Arg Pro Trp Asn Pro Pro Thr Phe Ser Pro
20 25 30
Ala Leu Leu Val Val Thr Glu Gly Asp Asn Ala Thr Phe Thr Cys Ser
35 40 45
Phe Ser Asn Thr Ser Glu Ser Phe Val Leu Asn Trp Tyr Arg Met Ser
50 55 60
Pro Ser Asn Gln Thr Asp Lys Leu Ala Ala Phe Pro Glu Asp Arg Ser
65 70 75 80
Gln Pro Gly Gln Asp Cys Arg Phe Arg Val Thr Gln Leu Pro Asn Gly
85 90 95
Arg Asp Phe His Met Ser Val Val Arg Ala Arg Arg Asn Asp Ser Gly
100 105 110
Thr Tyr Leu Cys Gly Ala Ile Ser Leu Ala Pro Lys Ala Gln Ile Lys
115 120 125
Glu Ser Leu Arg Ala Glu Leu Arg Val Thr Glu Arg Arg Ala Glu Val
130 135 140
Pro Thr Ala His Pro Ser Pro Ser Pro Arg Pro Ala Gly Gln Phe Gln
145 150 155 160
Gly Ser His His His His His His
165
<210> 24
<211> 145
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 24
Met Ser Ala Leu Leu Ile Leu Ala Leu Val Gly Ala Ala Val Val Trp
1 5 10 15
Ala Glu Ile Asn Gly Ser Ala Asn Tyr Glu Met Phe Ile Phe His Asn
20 25 30
Gly Gly Val Gln Ile Leu Cys Lys Tyr Pro Asp Ile Val Gln Gln Phe
35 40 45
Lys Met Gln Leu Leu Lys Gly Gly Gln Ile Leu Cys Asp Leu Thr Lys
50 55 60
Thr Lys Gly Ser Gly Asn Thr Val Ser Ile Lys Ser Leu Lys Phe Cys
65 70 75 80
His Ser Gln Leu Ser Asn Asn Ser Val Ser Phe Phe Leu Tyr Asn Leu
85 90 95
Asp His Ser His Ala Asn Tyr Tyr Phe Cys Asn Leu Ser Ile Phe Asp
100 105 110
Pro Pro Pro Phe Lys Val Thr Leu Thr Gly Gly Tyr Leu His Ile Tyr
115 120 125
Glu Ser Gln Leu Cys Cys Gln Leu Lys Gly Ser His His His His His
130 135 140
His
145
<210> 25
<211> 368
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 25
Met Ser Ala Leu Leu Ile Leu Ala Leu Val Gly Ala Ala Val Val Trp
1 5 10 15
Ala Gly Pro Phe Gln Gln Arg Gly Leu Phe Asp Phe Met Leu Glu Asp
20 25 30
Glu Ala Ser Gly Ile Gly Pro Glu Val Pro Asp Asp Arg Asp Phe Glu
35 40 45
Pro Ser Leu Gly Pro Val Cys Pro Phe Arg Cys Gln Cys His Leu Arg
50 55 60
Val Val Gln Cys Ser Asp Leu Gly Leu Asp Lys Val Pro Lys Asp Leu
65 70 75 80
Pro Pro Asp Thr Thr Leu Leu Asp Leu Gln Asn Asn Lys Ile Thr Glu
85 90 95
Ile Lys Asp Gly Asp Phe Lys Asn Leu Lys Asn Leu His Ala Leu Ile
100 105 110
Leu Val Asn Asn Lys Ile Ser Lys Val Ser Pro Gly Ala Phe Thr Pro
115 120 125
Leu Val Lys Leu Glu Arg Leu Tyr Leu Ser Lys Asn Gln Leu Lys Glu
130 135 140
Leu Pro Glu Lys Met Pro Lys Thr Leu Gln Glu Leu Arg Ala His Glu
145 150 155 160
Asn Glu Ile Thr Lys Val Arg Lys Val Thr Phe Asn Gly Leu Asn Gln
165 170 175
Met Ile Val Ile Glu Leu Gly Thr Asn Pro Leu Lys Ser Ser Gly Ile
180 185 190
Glu Asn Gly Ala Phe Gln Gly Met Lys Lys Leu Ser Tyr Ile Arg Ile
195 200 205
Ala Asp Thr Asn Ile Thr Ser Ile Pro Gln Gly Leu Pro Pro Ser Leu
210 215 220
Thr Glu Leu His Leu Asp Gly Asn Lys Ile Ser Arg Val Asp Ala Ala
225 230 235 240
Ser Leu Lys Gly Leu Asn Asn Leu Ala Lys Leu Gly Leu Ser Phe Asn
245 250 255
Ser Ile Ser Ala Val Asp Asn Gly Ser Leu Ala Asn Thr Pro His Leu
260 265 270
Arg Glu Leu His Leu Asp Asn Asn Lys Leu Thr Arg Val Pro Gly Gly
275 280 285
Leu Ala Glu His Lys Tyr Ile Gln Val Val Tyr Leu His Asn Asn Asn
290 295 300
Ile Ser Val Val Gly Ser Ser Asp Phe Cys Pro Pro Gly His Asn Thr
305 310 315 320
Lys Lys Ala Ser Tyr Ser Gly Val Ser Leu Phe Ser Asn Pro Val Gln
325 330 335
Tyr Trp Glu Ile Gln Pro Ser Thr Phe Arg Cys Val Tyr Val Arg Ser
340 345 350
Ala Ile Gln Leu Gly Asn Tyr Lys Gly Ser His His His His His His
355 360 365
<210> 26
<211> 267
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 26
Met Ser Ala Leu Leu Ile Leu Ala Leu Val Gly Ala Ala Val Val Trp
1 5 10 15
Ala Phe Val Arg Thr Lys Ile Asp Thr Thr Glu Asn Leu Leu Asn Thr
20 25 30
Glu Val His Ser Ser Pro Ala Gln Arg Trp Ser Met Gln Val Pro Pro
35 40 45
Glu Val Ser Ala Glu Ala Gly Asp Ala Ala Val Leu Pro Cys Thr Phe
50 55 60
Thr His Pro His Arg His Tyr Asp Gly Pro Leu Thr Ala Ile Trp Arg
65 70 75 80
Ala Gly Glu Pro Tyr Ala Gly Pro Gln Val Phe Arg Cys Ala Ala Ala
85 90 95
Arg Gly Ser Glu Leu Cys Gln Thr Ala Leu Ser His Gly Arg Phe Arg
100 105 110
Leu Leu Gly Asn Pro Arg Arg Asn Asp Leu Ser Leu Arg Val Glu Arg
115 120 125
Leu Ala Leu Ala Asp Asp Arg Arg Tyr Phe Cys Arg Val Glu Phe Ala
130 135 140
Gly Asp Val His Asp Arg Tyr Glu Ser Arg His Gly Val Arg Leu His
145 150 155 160
Val Thr Ala Ala Pro Arg Ile Val Asn Ile Ser Val Leu Pro Ser Pro
165 170 175
Ala His Ala Phe Arg Ala Leu Cys Thr Ala Glu Gly Glu Pro Pro Pro
180 185 190
Ala Leu Ala Trp Ser Gly Pro Ala Leu Gly Asn Ser Leu Ala Ala Val
195 200 205
Arg Ser Pro Arg Glu Gly His Gly His Leu Val Thr Ala Glu Leu Pro
210 215 220
Ala Leu Thr His Asp Gly Arg Tyr Thr Cys Thr Ala Ala Asn Ser Leu
225 230 235 240
Arg Ser Glu Ala Ser Val Tyr Leu Phe Arg Phe His Gly Ala Ser Gly
245 250 255
Ala Ser Thr Gly Ser His His His His His His
260 265
<210> 27
<211> 177
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 27
Met Ser Ala Leu Leu Ile Leu Ala Leu Val Gly Ala Ala Val Val Trp
1 5 10 15
Ala Asp Cys Asp Ile Glu Gly Lys Asp Gly Lys Gln Tyr Glu Ser Val
20 25 30
Leu Met Val Ser Ile Asp Gln Leu Leu Asp Ser Met Lys Glu Ile Gly
35 40 45
Ser Asn Cys Leu Asn Asn Glu Phe Asn Phe Phe Lys Arg His Ile Cys
50 55 60
Asp Ala Asn Lys Glu Gly Met Phe Leu Phe Arg Ala Ala Arg Lys Leu
65 70 75 80
Arg Gln Phe Leu Lys Met Asn Ser Thr Gly Asp Phe Asp Leu His Leu
85 90 95
Leu Lys Val Ser Glu Gly Thr Thr Ile Leu Leu Asn Cys Thr Gly Gln
100 105 110
Val Lys Gly Arg Lys Pro Ala Ala Leu Gly Glu Ala Gln Pro Thr Lys
115 120 125
Ser Leu Glu Glu Asn Lys Ser Leu Lys Glu Gln Lys Lys Leu Asn Asp
130 135 140
Leu Cys Phe Leu Lys Arg Leu Leu Gln Glu Ile Lys Thr Cys Trp Asn
145 150 155 160
Lys Ile Leu Met Gly Thr Lys Glu His Gly Ser His His His His His
165 170 175
His
<210> 28
<211> 158
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 28
Met Ser Ala Leu Leu Ile Leu Ala Leu Val Gly Ala Ala Val Val Trp
1 5 10 15
Ala Arg Lys Ile Pro Lys Val Gly His Thr Phe Phe Gln Lys Pro Glu
20 25 30
Ser Cys Pro Pro Val Pro Gly Gly Ser Met Lys Leu Asp Ile Gly Ile
35 40 45
Ile Asn Glu Asn Gln Arg Val Ser Met Ser Arg Asn Ile Glu Ser Arg
50 55 60
Ser Thr Ser Pro Trp Asn Tyr Thr Val Thr Trp Asp Pro Asn Arg Tyr
65 70 75 80
Pro Ser Glu Val Val Gln Ala Gln Cys Arg Asn Leu Gly Cys Ile Asn
85 90 95
Ala Gln Gly Lys Glu Asp Ile Ser Met Asn Ser Val Pro Ile Gln Gln
100 105 110
Glu Thr Leu Val Val Arg Arg Lys His Gln Gly Cys Ser Val Ser Phe
115 120 125
Gln Leu Glu Lys Val Leu Val Thr Val Gly Cys Thr Cys Val Thr Pro
130 135 140
Val Ile His His Val Gln Gly Ser His His His His His His
145 150 155
<210> 29
<211> 547
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 29
Met Ser Ala Leu Leu Ile Leu Ala Leu Val Gly Ala Ala Val Val Trp
1 5 10 15
Ala Trp Glu Leu Thr Ile Leu His Thr Asn Asp Val His Ser Arg Leu
20 25 30
Glu Gln Thr Ser Glu Asp Ser Ser Lys Cys Val Asn Ala Ser Arg Cys
35 40 45
Met Gly Gly Val Ala Arg Leu Phe Thr Lys Val Gln Gln Ile Arg Arg
50 55 60
Ala Glu Pro Asn Val Leu Leu Leu Asp Ala Gly Asp Gln Tyr Gln Gly
65 70 75 80
Thr Ile Trp Phe Thr Val Tyr Lys Gly Ala Glu Val Ala His Phe Met
85 90 95
Asn Ala Leu Arg Tyr Asp Ala Met Ala Leu Gly Asn His Glu Phe Asp
100 105 110
Asn Gly Val Glu Gly Leu Ile Glu Pro Leu Leu Lys Glu Ala Lys Phe
115 120 125
Pro Ile Leu Ser Ala Asn Ile Lys Ala Lys Gly Pro Ala Ser Gln Ile
130 135 140
Ser Gly Leu Tyr Leu Pro Tyr Lys Val Leu Pro Val Gly Asp Glu Val
145 150 155 160
Val Gly Ile Val Gly Tyr Thr Ser Lys Glu Thr Pro Phe Leu Ser Asn
165 170 175
Pro Gly Thr Asn Leu Val Phe Glu Asp Glu Ile Thr Ala Leu Gln Pro
180 185 190
Glu Val Asp Lys Leu Lys Thr Leu Asn Val Asn Lys Ile Ile Ala Leu
195 200 205
Gly His Ser Gly Phe Glu Met Asp Lys Leu Ile Ala Gln Lys Val Arg
210 215 220
Gly Val Asp Val Val Val Gly Gly His Ser Asn Thr Phe Leu Tyr Thr
225 230 235 240
Gly Asn Pro Pro Ser Lys Glu Val Pro Ala Gly Lys Tyr Pro Phe Ile
245 250 255
Val Thr Ser Asp Asp Gly Arg Lys Val Pro Val Val Gln Ala Tyr Ala
260 265 270
Phe Gly Lys Tyr Leu Gly Tyr Leu Lys Ile Glu Phe Asp Glu Arg Gly
275 280 285
Asn Val Ile Ser Ser His Gly Asn Pro Ile Leu Leu Asn Ser Ser Ile
290 295 300
Pro Glu Asp Pro Ser Ile Lys Ala Asp Ile Asn Lys Trp Arg Ile Lys
305 310 315 320
Leu Asp Asn Tyr Ser Thr Gln Glu Leu Gly Lys Thr Ile Val Tyr Leu
325 330 335
Asp Gly Ser Ser Gln Ser Cys Arg Phe Arg Glu Cys Asn Met Gly Asn
340 345 350
Leu Ile Cys Asp Ala Met Ile Asn Asn Asn Leu Arg His Thr Asp Glu
355 360 365
Met Phe Trp Asn His Val Ser Met Cys Ile Leu Asn Gly Gly Gly Ile
370 375 380
Arg Ser Pro Ile Asp Glu Arg Asn Asn Gly Thr Ile Thr Trp Glu Asn
385 390 395 400
Leu Ala Ala Val Leu Pro Phe Gly Gly Thr Phe Asp Leu Val Gln Leu
405 410 415
Lys Gly Ser Thr Leu Lys Lys Ala Phe Glu His Ser Val His Arg Tyr
420 425 430
Gly Gln Ser Thr Gly Glu Phe Leu Gln Val Gly Gly Ile His Val Val
435 440 445
Tyr Asp Leu Ser Arg Lys Pro Gly Asp Arg Val Val Lys Leu Asp Val
450 455 460
Leu Cys Thr Lys Cys Arg Val Pro Ser Tyr Asp Pro Leu Lys Met Asp
465 470 475 480
Glu Val Tyr Lys Val Ile Leu Pro Asn Phe Leu Ala Asn Gly Gly Asp
485 490 495
Gly Phe Gln Met Ile Lys Asp Glu Leu Leu Arg His Asp Ser Gly Asp
500 505 510
Gln Asp Ile Asn Val Val Ser Thr Tyr Ile Ser Lys Met Lys Val Ile
515 520 525
Tyr Pro Ala Val Glu Gly Arg Ile Lys Phe Ser Gly Ser His His His
530 535 540
His His His
545
<210> 30
<211> 473
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 30
Met Pro Val Pro Trp Phe Leu Leu Ser Leu Ala Leu Gly Arg Ser Pro
1 5 10 15
Val Val Leu Ser Leu Glu Arg Leu Val Gly Pro Gln Asp Ala Thr His
20 25 30
Cys Ser Pro Gly Leu Ser Cys Arg Leu Trp Asp Ser Asp Ile Leu Cys
35 40 45
Leu Pro Gly Asp Ile Val Pro Ala Pro Gly Pro Val Leu Ala Pro Thr
50 55 60
His Leu Gln Thr Glu Leu Val Leu Arg Cys Gln Lys Glu Thr Asp Cys
65 70 75 80
Asp Leu Cys Leu Arg Val Ala Val His Leu Ala Val His Gly His Trp
85 90 95
Glu Glu Pro Glu Asp Glu Glu Lys Phe Gly Gly Ala Ala Asp Ser Gly
100 105 110
Val Glu Glu Pro Arg Asn Ala Ser Leu Gln Ala Gln Val Val Leu Ser
115 120 125
Phe Gln Ala Tyr Pro Thr Ala Arg Cys Val Leu Leu Glu Val Gln Val
130 135 140
Pro Ala Ala Leu Val Gln Phe Gly Gln Ser Val Gly Ser Val Val Tyr
145 150 155 160
Asp Cys Phe Glu Ala Ala Leu Gly Ser Glu Val Arg Ile Trp Ser Tyr
165 170 175
Thr Gln Pro Arg Tyr Glu Lys Glu Leu Asn His Thr Gln Gln Leu Pro
180 185 190
Asp Cys Arg Gly Leu Glu Val Trp Asn Ser Ile Pro Ser Cys Trp Ala
195 200 205
Leu Pro Trp Leu Asn Val Ser Ala Asp Gly Asp Asn Val His Leu Val
210 215 220
Leu Asn Val Ser Glu Glu Gln His Phe Gly Leu Ser Leu Tyr Trp Asn
225 230 235 240
Gln Val Gln Gly Pro Pro Lys Pro Arg Trp His Lys Asn Leu Thr Gly
245 250 255
Pro Gln Ile Ile Thr Leu Asn His Thr Asp Leu Val Pro Cys Leu Cys
260 265 270
Ile Gln Val Trp Pro Leu Glu Pro Asp Ser Val Arg Thr Asn Ile Cys
275 280 285
Pro Phe Arg Glu Asp Pro Arg Ala His Gln Asn Leu Trp Gln Ala Ala
290 295 300
Arg Leu Gln Leu Leu Thr Leu Gln Ser Trp Leu Leu Asp Ala Pro Cys
305 310 315 320
Ser Leu Pro Ala Glu Ala Ala Leu Cys Trp Arg Ala Pro Gly Gly Asp
325 330 335
Pro Cys Gln Pro Leu Val Pro Pro Leu Ser Trp Glu Asn Val Thr Val
340 345 350
Asp Lys Val Leu Glu Phe Pro Leu Leu Lys Gly His Pro Asn Leu Cys
355 360 365
Val Gln Val Asn Ser Ser Glu Lys Leu Gln Leu Gln Glu Cys Leu Trp
370 375 380
Ala Asp Ser Leu Gly Pro Leu Lys Asp Asp Val Leu Leu Leu Glu Thr
385 390 395 400
Arg Gly Pro Gln Asp Asn Arg Ser Leu Cys Ala Leu Glu Pro Ser Gly
405 410 415
Cys Thr Ser Leu Pro Ser Lys Ala Ser Thr Arg Ala Ala Arg Leu Gly
420 425 430
Glu Tyr Leu Leu Gln Asp Leu Gln Ser Gly Gln Cys Leu Gln Leu Trp
435 440 445
Asp Asp Asp Leu Gly Ala Leu Trp Ala Cys Pro Met Asp Lys Tyr Ile
450 455 460
His Gly Ser His His His His His His
465 470
<210> 31
<211> 160
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 31
Met Leu Arg Leu Leu Leu Ala Leu Asn Leu Phe Pro Ser Ile Gln Val
1 5 10 15
Thr Gly Asn Lys Ile Leu Val Lys Gln Ser Pro Met Leu Val Ala Tyr
20 25 30
Asp Asn Ala Val Asn Leu Ser Cys Lys Tyr Ser Tyr Asn Leu Phe Ser
35 40 45
Arg Glu Phe Arg Ala Ser Leu His Lys Gly Leu Asp Ser Ala Val Glu
50 55 60
Val Cys Val Val Tyr Gly Asn Tyr Ser Gln Gln Leu Gln Val Tyr Ser
65 70 75 80
Lys Thr Gly Phe Asn Cys Asp Gly Lys Leu Gly Asn Glu Ser Val Thr
85 90 95
Phe Tyr Leu Gln Asn Leu Tyr Val Asn Gln Thr Asp Ile Tyr Phe Cys
100 105 110
Lys Ile Glu Val Met Tyr Pro Pro Pro Tyr Leu Asp Asn Glu Lys Ser
115 120 125
Asn Gly Thr Ile Ile His Val Lys Gly Lys His Leu Cys Pro Ser Pro
130 135 140
Leu Phe Pro Gly Pro Ser Lys Pro Gly Ser His His His His His His
145 150 155 160
<210> 32
<211> 175
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 32
Met Gln Ile Pro Gln Ala Pro Trp Pro Val Val Trp Ala Val Leu Gln
1 5 10 15
Leu Gly Trp Arg Pro Gly Trp Phe Leu Asp Ser Pro Asp Arg Pro Trp
20 25 30
Asn Pro Pro Thr Phe Ser Pro Ala Leu Leu Val Val Thr Glu Gly Asp
35 40 45
Asn Ala Thr Phe Thr Cys Ser Phe Ser Asn Thr Ser Glu Ser Phe Val
50 55 60
Leu Asn Trp Tyr Arg Met Ser Pro Ser Asn Gln Thr Asp Lys Leu Ala
65 70 75 80
Ala Phe Pro Glu Asp Arg Ser Gln Pro Gly Gln Asp Cys Arg Phe Arg
85 90 95
Val Thr Gln Leu Pro Asn Gly Arg Asp Phe His Met Ser Val Val Arg
100 105 110
Ala Arg Arg Asn Asp Ser Gly Thr Tyr Leu Cys Gly Ala Ile Ser Leu
115 120 125
Ala Pro Lys Ala Gln Ile Lys Glu Ser Leu Arg Ala Glu Leu Arg Val
130 135 140
Thr Glu Arg Arg Ala Glu Val Pro Thr Ala His Pro Ser Pro Ser Pro
145 150 155 160
Arg Pro Ala Gly Gln Phe Gln Gly Ser His His His His His His
165 170 175
<210> 33
<211> 148
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 33
Met Lys Ser Gly Leu Trp Tyr Phe Phe Leu Phe Cys Leu Arg Ile Lys
1 5 10 15
Val Leu Thr Gly Glu Ile Asn Gly Ser Ala Asn Tyr Glu Met Phe Ile
20 25 30
Phe His Asn Gly Gly Val Gln Ile Leu Cys Lys Tyr Pro Asp Ile Val
35 40 45
Gln Gln Phe Lys Met Gln Leu Leu Lys Gly Gly Gln Ile Leu Cys Asp
50 55 60
Leu Thr Lys Thr Lys Gly Ser Gly Asn Thr Val Ser Ile Lys Ser Leu
65 70 75 80
Lys Phe Cys His Ser Gln Leu Ser Asn Asn Ser Val Ser Phe Phe Leu
85 90 95
Tyr Asn Leu Asp His Ser His Ala Asn Tyr Tyr Phe Cys Asn Leu Ser
100 105 110
Ile Phe Asp Pro Pro Pro Phe Lys Val Thr Leu Thr Gly Gly Tyr Leu
115 120 125
His Ile Tyr Glu Ser Gln Leu Cys Cys Gln Leu Lys Gly Ser His His
130 135 140
His His His His
145
<210> 34
<211> 367
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 34
Met Lys Ala Thr Ile Ile Leu Leu Leu Leu Ala Gln Val Ser Trp Ala
1 5 10 15
Gly Pro Phe Gln Gln Arg Gly Leu Phe Asp Phe Met Leu Glu Asp Glu
20 25 30
Ala Ser Gly Ile Gly Pro Glu Val Pro Asp Asp Arg Asp Phe Glu Pro
35 40 45
Ser Leu Gly Pro Val Cys Pro Phe Arg Cys Gln Cys His Leu Arg Val
50 55 60
Val Gln Cys Ser Asp Leu Gly Leu Asp Lys Val Pro Lys Asp Leu Pro
65 70 75 80
Pro Asp Thr Thr Leu Leu Asp Leu Gln Asn Asn Lys Ile Thr Glu Ile
85 90 95
Lys Asp Gly Asp Phe Lys Asn Leu Lys Asn Leu His Ala Leu Ile Leu
100 105 110
Val Asn Asn Lys Ile Ser Lys Val Ser Pro Gly Ala Phe Thr Pro Leu
115 120 125
Val Lys Leu Glu Arg Leu Tyr Leu Ser Lys Asn Gln Leu Lys Glu Leu
130 135 140
Pro Glu Lys Met Pro Lys Thr Leu Gln Glu Leu Arg Ala His Glu Asn
145 150 155 160
Glu Ile Thr Lys Val Arg Lys Val Thr Phe Asn Gly Leu Asn Gln Met
165 170 175
Ile Val Ile Glu Leu Gly Thr Asn Pro Leu Lys Ser Ser Gly Ile Glu
180 185 190
Asn Gly Ala Phe Gln Gly Met Lys Lys Leu Ser Tyr Ile Arg Ile Ala
195 200 205
Asp Thr Asn Ile Thr Ser Ile Pro Gln Gly Leu Pro Pro Ser Leu Thr
210 215 220
Glu Leu His Leu Asp Gly Asn Lys Ile Ser Arg Val Asp Ala Ala Ser
225 230 235 240
Leu Lys Gly Leu Asn Asn Leu Ala Lys Leu Gly Leu Ser Phe Asn Ser
245 250 255
Ile Ser Ala Val Asp Asn Gly Ser Leu Ala Asn Thr Pro His Leu Arg
260 265 270
Glu Leu His Leu Asp Asn Asn Lys Leu Thr Arg Val Pro Gly Gly Leu
275 280 285
Ala Glu His Lys Tyr Ile Gln Val Val Tyr Leu His Asn Asn Asn Ile
290 295 300
Ser Val Val Gly Ser Ser Asp Phe Cys Pro Pro Gly His Asn Thr Lys
305 310 315 320
Lys Ala Ser Tyr Ser Gly Val Ser Leu Phe Ser Asn Pro Val Gln Tyr
325 330 335
Trp Glu Ile Gln Pro Ser Thr Phe Arg Cys Val Tyr Val Arg Ser Ala
340 345 350
Ile Gln Leu Gly Asn Tyr Lys Gly Ser His His His His His His
355 360 365
<210> 35
<211> 269
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 35
Met Glu Lys Ser Ile Trp Leu Leu Ala Cys Leu Ala Trp Val Leu Pro
1 5 10 15
Thr Gly Ser Phe Val Arg Thr Lys Ile Asp Thr Thr Glu Asn Leu Leu
20 25 30
Asn Thr Glu Val His Ser Ser Pro Ala Gln Arg Trp Ser Met Gln Val
35 40 45
Pro Pro Glu Val Ser Ala Glu Ala Gly Asp Ala Ala Val Leu Pro Cys
50 55 60
Thr Phe Thr His Pro His Arg His Tyr Asp Gly Pro Leu Thr Ala Ile
65 70 75 80
Trp Arg Ala Gly Glu Pro Tyr Ala Gly Pro Gln Val Phe Arg Cys Ala
85 90 95
Ala Ala Arg Gly Ser Glu Leu Cys Gln Thr Ala Leu Ser His Gly Arg
100 105 110
Phe Arg Leu Leu Gly Asn Pro Arg Arg Asn Asp Leu Ser Leu Arg Val
115 120 125
Glu Arg Leu Ala Leu Ala Asp Asp Arg Arg Tyr Phe Cys Arg Val Glu
130 135 140
Phe Ala Gly Asp Val His Asp Arg Tyr Glu Ser Arg His Gly Val Arg
145 150 155 160
Leu His Val Thr Ala Ala Pro Arg Ile Val Asn Ile Ser Val Leu Pro
165 170 175
Ser Pro Ala His Ala Phe Arg Ala Leu Cys Thr Ala Glu Gly Glu Pro
180 185 190
Pro Pro Ala Leu Ala Trp Ser Gly Pro Ala Leu Gly Asn Ser Leu Ala
195 200 205
Ala Val Arg Ser Pro Arg Glu Gly His Gly His Leu Val Thr Ala Glu
210 215 220
Leu Pro Ala Leu Thr His Asp Gly Arg Tyr Thr Cys Thr Ala Ala Asn
225 230 235 240
Ser Leu Arg Ser Glu Ala Ser Val Tyr Leu Phe Arg Phe His Gly Ala
245 250 255
Ser Gly Ala Ser Thr Gly Ser His His His His His His
260 265
<210> 36
<211> 185
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 36
Met Phe His Val Ser Phe Arg Tyr Ile Phe Gly Leu Pro Pro Leu Ile
1 5 10 15
Leu Val Leu Leu Pro Val Ala Ser Ser Asp Cys Asp Ile Glu Gly Lys
20 25 30
Asp Gly Lys Gln Tyr Glu Ser Val Leu Met Val Ser Ile Asp Gln Leu
35 40 45
Leu Asp Ser Met Lys Glu Ile Gly Ser Asn Cys Leu Asn Asn Glu Phe
50 55 60
Asn Phe Phe Lys Arg His Ile Cys Asp Ala Asn Lys Glu Gly Met Phe
65 70 75 80
Leu Phe Arg Ala Ala Arg Lys Leu Arg Gln Phe Leu Lys Met Asn Ser
85 90 95
Thr Gly Asp Phe Asp Leu His Leu Leu Lys Val Ser Glu Gly Thr Thr
100 105 110
Ile Leu Leu Asn Cys Thr Gly Gln Val Lys Gly Arg Lys Pro Ala Ala
115 120 125
Leu Gly Glu Ala Gln Pro Thr Lys Ser Leu Glu Glu Asn Lys Ser Leu
130 135 140
Lys Glu Gln Lys Lys Leu Asn Asp Leu Cys Phe Leu Lys Arg Leu Leu
145 150 155 160
Gln Glu Ile Lys Thr Cys Trp Asn Lys Ile Leu Met Gly Thr Lys Glu
165 170 175
His Gly Ser His His His His His His
180 185
<210> 37
<211> 171
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 37
Met Thr Val Lys Thr Leu His Gly Pro Ala Met Val Lys Tyr Leu Leu
1 5 10 15
Leu Ser Ile Leu Gly Leu Ala Phe Leu Ser Glu Ala Ala Ala Arg Lys
20 25 30
Ile Pro Lys Val Gly His Thr Phe Phe Gln Lys Pro Glu Ser Cys Pro
35 40 45
Pro Val Pro Gly Gly Ser Met Lys Leu Asp Ile Gly Ile Ile Asn Glu
50 55 60
Asn Gln Arg Val Ser Met Ser Arg Asn Ile Glu Ser Arg Ser Thr Ser
65 70 75 80
Pro Trp Asn Tyr Thr Val Thr Trp Asp Pro Asn Arg Tyr Pro Ser Glu
85 90 95
Val Val Gln Ala Gln Cys Arg Asn Leu Gly Cys Ile Asn Ala Gln Gly
100 105 110
Lys Glu Asp Ile Ser Met Asn Ser Val Pro Ile Gln Gln Glu Thr Leu
115 120 125
Val Val Arg Arg Lys His Gln Gly Cys Ser Val Ser Phe Gln Leu Glu
130 135 140
Lys Val Leu Val Thr Val Gly Cys Thr Cys Val Thr Pro Val Ile His
145 150 155 160
His Val Gln Gly Ser His His His His His His
165 170
<210> 38
<211> 556
<212> PRT
<213> Artificial Sequence
<220>
<223> recombinant protein
<400> 38
Met Cys Pro Arg Ala Ala Arg Ala Pro Ala Thr Leu Leu Leu Ala Leu
1 5 10 15
Gly Ala Val Leu Trp Pro Ala Ala Gly Ala Trp Glu Leu Thr Ile Leu
20 25 30
His Thr Asn Asp Val His Ser Arg Leu Glu Gln Thr Ser Glu Asp Ser
35 40 45
Ser Lys Cys Val Asn Ala Ser Arg Cys Met Gly Gly Val Ala Arg Leu
50 55 60
Phe Thr Lys Val Gln Gln Ile Arg Arg Ala Glu Pro Asn Val Leu Leu
65 70 75 80
Leu Asp Ala Gly Asp Gln Tyr Gln Gly Thr Ile Trp Phe Thr Val Tyr
85 90 95
Lys Gly Ala Glu Val Ala His Phe Met Asn Ala Leu Arg Tyr Asp Ala
100 105 110
Met Ala Leu Gly Asn His Glu Phe Asp Asn Gly Val Glu Gly Leu Ile
115 120 125
Glu Pro Leu Leu Lys Glu Ala Lys Phe Pro Ile Leu Ser Ala Asn Ile
130 135 140
Lys Ala Lys Gly Pro Ala Ser Gln Ile Ser Gly Leu Tyr Leu Pro Tyr
145 150 155 160
Lys Val Leu Pro Val Gly Asp Glu Val Val Gly Ile Val Gly Tyr Thr
165 170 175
Ser Lys Glu Thr Pro Phe Leu Ser Asn Pro Gly Thr Asn Leu Val Phe
180 185 190
Glu Asp Glu Ile Thr Ala Leu Gln Pro Glu Val Asp Lys Leu Lys Thr
195 200 205
Leu Asn Val Asn Lys Ile Ile Ala Leu Gly His Ser Gly Phe Glu Met
210 215 220
Asp Lys Leu Ile Ala Gln Lys Val Arg Gly Val Asp Val Val Val Gly
225 230 235 240
Gly His Ser Asn Thr Phe Leu Tyr Thr Gly Asn Pro Pro Ser Lys Glu
245 250 255
Val Pro Ala Gly Lys Tyr Pro Phe Ile Val Thr Ser Asp Asp Gly Arg
260 265 270
Lys Val Pro Val Val Gln Ala Tyr Ala Phe Gly Lys Tyr Leu Gly Tyr
275 280 285
Leu Lys Ile Glu Phe Asp Glu Arg Gly Asn Val Ile Ser Ser His Gly
290 295 300
Asn Pro Ile Leu Leu Asn Ser Ser Ile Pro Glu Asp Pro Ser Ile Lys
305 310 315 320
Ala Asp Ile Asn Lys Trp Arg Ile Lys Leu Asp Asn Tyr Ser Thr Gln
325 330 335
Glu Leu Gly Lys Thr Ile Val Tyr Leu Asp Gly Ser Ser Gln Ser Cys
340 345 350
Arg Phe Arg Glu Cys Asn Met Gly Asn Leu Ile Cys Asp Ala Met Ile
355 360 365
Asn Asn Asn Leu Arg His Thr Asp Glu Met Phe Trp Asn His Val Ser
370 375 380
Met Cys Ile Leu Asn Gly Gly Gly Ile Arg Ser Pro Ile Asp Glu Arg
385 390 395 400
Asn Asn Gly Thr Ile Thr Trp Glu Asn Leu Ala Ala Val Leu Pro Phe
405 410 415
Gly Gly Thr Phe Asp Leu Val Gln Leu Lys Gly Ser Thr Leu Lys Lys
420 425 430
Ala Phe Glu His Ser Val His Arg Tyr Gly Gln Ser Thr Gly Glu Phe
435 440 445
Leu Gln Val Gly Gly Ile His Val Val Tyr Asp Leu Ser Arg Lys Pro
450 455 460
Gly Asp Arg Val Val Lys Leu Asp Val Leu Cys Thr Lys Cys Arg Val
465 470 475 480
Pro Ser Tyr Asp Pro Leu Lys Met Asp Glu Val Tyr Lys Val Ile Leu
485 490 495
Pro Asn Phe Leu Ala Asn Gly Gly Asp Gly Phe Gln Met Ile Lys Asp
500 505 510
Glu Leu Leu Arg His Asp Ser Gly Asp Gln Asp Ile Asn Val Val Ser
515 520 525
Thr Tyr Ile Ser Lys Met Lys Val Ile Tyr Pro Ala Val Glu Gly Arg
530 535 540
Ile Lys Phe Ser Gly Ser His His His His His His
545 550 555
<210> 39
<211> 1413
<212> DNA
<213> Artificial Sequence
<220>
<223> sequence encoding Signal peptide + Human IL-17RC + poly His tag of the present invention
<400> 39
atgagcgccc tgctgatcct ggccctggtg ggcgccgccg tggtgtgggc cctggagcgc 60
ctggtgggcc cccaggacgc cacccactgc agccccggcc tgagctgccg cctgtgggac 120
agcgacatcc tgtgcctgcc cggcgacatc gtgcccgccc ccggccccgt gctggccccc 180
acccacctgc agaccgagct ggtgctgcgc tgccagaagg agaccgactg cgacctgtgc 240
ctgcgcgtgg ccgtgcacct ggccgtgcac ggccactggg aggagcccga ggacgaggag 300
aagttcggcg gcgccgccga cagcggcgtg gaggagcccc gcaacgccag cctgcaggcc 360
caggtggtgc tgagcttcca ggcctacccc accgcccgct gcgtgctgct ggaggtgcag 420
gtgcccgccg ccctggtgca gttcggccag agcgtgggca gcgtggtgta cgactgcttc 480
gaggccgccc tgggcagcga ggtgcgcatc tggagctaca cccagccccg ctacgagaag 540
gagctgaacc acacccagca gctgcccgac tgccgcggcc tggaggtgtg gaacagcatc 600
cccagctgct gggccctgcc ctggctgaac gtgagcgccg acggcgacaa cgtgcacctg 660
gtgctgaacg tgagcgagga gcagcacttc ggcctgagcc tgtactggaa ccaggtgcag 720
ggccccccca agccccgctg gcacaagaac ctgaccggcc cccagatcat caccctgaac 780
cacaccgacc tggtgccctg cctgtgcatc caggtgtggc ccctggagcc cgacagcgtg 840
cgcaccaaca tctgcccctt ccgcgaggac ccccgcgccc accagaacct gtggcaggcc 900
gcccgcctgc agctgctgac cctgcagagc tggctgctgg acgccccctg cagcctgccc 960
gccgaggccg ccctgtgctg gcgcgccccc ggcggcgacc cctgccagcc cctggtgccc 1020
cccctgagct gggagaacgt gaccgtggac aaggtgctgg agttccccct gctgaagggc 1080
caccccaacc tgtgcgtgca ggtgaacagc agcgagaagc tgcagctgca ggagtgcctg 1140
tgggccgaca gcctgggccc cctgaaggac gacgtgctgc tgctggagac ccgcggcccc 1200
caggacaacc gcagcctgtg cgccctggag cccagcggct gcaccagcct gcccagcaag 1260
gccagcaccc gcgccgcccg cctgggcgag tacctgctgc aggacctgca gagcggccag 1320
tgcctgcagc tgtgggacga cgacctgggc gccctgtggg cctgccccat ggacaagtac 1380
atccacggca gccaccacca ccaccaccac taa 1413
<210> 40
<211> 480
<212> DNA
<213> Artificial Sequence
<220>
<223> sequence encoding Signal peptide + Human CD28+ poly His tag of the present invention
<400> 40
atgagcgccc tgctgatcct ggccctggtg ggcgccgccg tggtgtgggc caacaagatc 60
ctggtgaagc agagccccat gctggtggcc tacgacaacg ccgtgaacct gagctgcaag 120
tacagctaca acctgttcag ccgcgagttc cgcgccagcc tgcacaaggg cctggacagc 180
gccgtggagg tgtgcgtggt gtacggcaac tacagccagc agctgcaggt gtacagcaag 240
accggcttca actgcgacgg caagctgggc aacgagagcg tgaccttcta cctgcagaac 300
ctgtacgtga accagaccga catctacttc tgcaagatcg aggtgatgta cccccccccc 360
tacctggaca acgagaagag caacggcacc atcatccacg tgaagggcaa gcacctgtgc 420
cccagccccc tgttccccgg ccccagcaag cccggcagcc accaccacca ccaccactaa 480
<210> 41
<211> 507
<212> DNA
<213> Artificial Sequence
<220>
<223> sequence encoding signal peptide + Human PD-1+ poly His tag of the present invention
<400> 41
atgagcgccc tgctgatcct ggccctggtg ggcgccgccg tggtgtgggc cctggacagc 60
cccgaccgcc cctggaaccc ccccaccttc agccccgccc tgctggtggt gaccgagggc 120
gacaacgcca ccttcacctg cagcttcagc aacaccagcg agagcttcgt gctgaactgg 180
taccgcatga gccccagcaa ccagaccgac aagctggccg ccttccccga ggaccgcagc 240
cagcccggcc aggactgccg cttccgcgtg acccagctgc ccaacggccg cgacttccac 300
atgagcgtgg tgcgcgcccg ccgcaacgac agcggcacct acctgtgcgg cgccatcagc 360
ctggccccca aggcccagat caaggagagc ctgcgcgccg agctgcgcgt gaccgagcgc 420
cgcgccgagg tgcccaccgc ccaccccagc cccagccccc gccccgccgg ccagttccag 480
ggcagccacc accaccacca ccactaa 507
<210> 42
<211> 438
<212> DNA
<213> Artificial Sequence
<220>
<223> sequence encoding Signal peptide + Human ICOS + poly His tag of the present invention
<400> 42
atgagcgccc tgctgatcct ggccctggtg ggcgccgccg tggtgtgggc cgagatcaac 60
ggcagcgcca actacgagat gttcatcttc cacaacggcg gcgtgcagat cctgtgcaag 120
taccccgaca tcgtgcagca gttcaagatg cagctgctga agggcggcca gatcctgtgc 180
gacctgacca agaccaaggg cagcggcaac accgtgagca tcaagagcct gaagttctgc 240
cacagccagc tgagcaacaa cagcgtgagc ttcttcctgt acaacctgga ccacagccac 300
gccaactact acttctgcaa cctgagcatc ttcgaccccc cccccttcaa ggtgaccctg 360
accggcggct acctgcacat ctacgagagc cagctgtgct gccagctgaa gggcagccac 420
caccaccacc accactaa 438
<210> 43
<211> 1107
<212> DNA
<213> Artificial Sequence
<220>
<223> sequence encoding Signal peptide + Human Decorin + Poly His tag of the present invention
<400> 43
atgagcgccc tgctgatcct ggccctggtg ggcgccgccg tggtgtgggc cggccccttc 60
cagcagcgcg gcctgttcga cttcatgctg gaggacgagg ccagcggcat cggccccgag 120
gtgcccgacg accgcgactt cgagcccagc ctgggccccg tgtgcccctt ccgctgccag 180
tgccacctgc gcgtggtgca gtgcagcgac ctgggcctgg acaaggtgcc caaggacctg 240
ccccccgaca ccaccctgct ggacctgcag aacaacaaga tcaccgagat caaggacggc 300
gacttcaaga acctgaagaa cctgcacgcc ctgatcctgg tgaacaacaa gatcagcaag 360
gtgagccccg gcgccttcac ccccctggtg aagctggagc gcctgtacct gagcaagaac 420
cagctgaagg agctgcccga gaagatgccc aagaccctgc aggagctgcg cgcccacgag 480
aacgagatca ccaaggtgcg caaggtgacc ttcaacggcc tgaaccagat gatcgtgatc 540
gagctgggca ccaaccccct gaagagcagc ggcatcgaga acggcgcctt ccagggcatg 600
aagaagctga gctacatccg catcgccgac accaacatca ccagcatccc ccagggcctg 660
ccccccagcc tgaccgagct gcacctggac ggcaacaaga tcagccgcgt ggacgccgcc 720
agcctgaagg gcctgaacaa cctggccaag ctgggcctga gcttcaacag catcagcgcc 780
gtggacaacg gcagcctggc caacaccccc cacctgcgcg agctgcacct ggacaacaac 840
aagctgaccc gcgtgcccgg cggcctggcc gagcacaagt acatccaggt ggtgtacctg 900
cacaacaaca acatcagcgt ggtgggcagc agcgacttct gcccccccgg ccacaacacc 960
aagaaggcca gctacagcgg cgtgagcctg ttcagcaacc ccgtgcagta ctgggagatc 1020
cagcccagca ccttccgctg cgtgtacgtg cgcagcgcca tccagctggg caactacaag 1080
ggcagccacc accaccacca ccactaa 1107
<210> 44
<211> 804
<212> DNA
<213> Artificial Sequence
<220>
<223> sequence encoding Signal peptide + Human Siglec-15+ poly His tag of the present invention
<400> 44
atgagcgccc tgctgatcct ggccctggtg ggcgccgccg tggtgtgggc cttcgtgcgc 60
accaagatcg acaccaccga gaacctgctg aacaccgagg tgcacagcag ccccgcccag 120
cgctggagca tgcaggtgcc ccccgaggtg agcgccgagg ccggcgacgc cgccgtgctg 180
ccctgcacct tcacccaccc ccaccgccac tacgacggcc ccctgaccgc catctggcgc 240
gccggcgagc cctacgccgg cccccaggtg ttccgctgcg ccgccgcccg cggcagcgag 300
ctgtgccaga ccgccctgag ccacggccgc ttccgcctgc tgggcaaccc ccgccgcaac 360
gacctgagcc tgcgcgtgga gcgcctggcc ctggccgacg accgccgcta cttctgccgc 420
gtggagttcg ccggcgacgt gcacgaccgc tacgagagcc gccacggcgt gcgcctgcac 480
gtgaccgccg ccccccgcat cgtgaacatc agcgtgctgc ccagccccgc ccacgccttc 540
cgcgccctgt gcaccgccga gggcgagccc ccccccgccc tggcctggag cggccccgcc 600
ctgggcaaca gcctggccgc cgtgcgcagc ccccgcgagg gccacggcca cctggtgacc 660
gccgagctgc ccgccctgac ccacgacggc cgctacacct gcaccgccgc caacagcctg 720
cgcagcgagg ccagcgtgta cctgttccgc ttccacggcg ccagcggcgc cagcaccggc 780
agccaccacc accaccacca ctaa 804
<210> 45
<211> 534
<212> DNA
<213> Artificial Sequence
<220>
<223> sequence encoding signal peptide + Human IL-7+ poly His tag of the present invention
<400> 45
atgagcgccc tgctgatcct ggccctggtg ggcgccgccg tggtgtgggc cgactgcgac 60
atcgagggca aggacggcaa gcagtacgag agcgtgctga tggtgagcat cgaccagctg 120
ctggacagca tgaaggagat cggcagcaac tgcctgaaca acgagttcaa cttcttcaag 180
cgccacatct gcgacgccaa caaggagggc atgttcctgt tccgcgccgc ccgcaagctg 240
cgccagttcc tgaagatgaa cagcaccggc gacttcgacc tgcacctgct gaaggtgagc 300
gagggcacca ccatcctgct gaactgcacc ggccaggtga agggccgcaa gcccgccgcc 360
ctgggcgagg cccagcccac caagagcctg gaggagaaca agagcctgaa ggagcagaag 420
aagctgaacg acctgtgctt cctgaagcgc ctgctgcagg agatcaagac ctgctggaac 480
aagatcctga tgggcaccaa ggagcacggc agccaccacc accaccacca ctaa 534
<210> 46
<211> 477
<212> DNA
<213> Artificial Sequence
<220>
<223> sequence encoding Signal peptide + HumanIL-17F + poly His tag of the present invention
<400> 46
atgagcgccc tgctgatcct ggccctggtg ggcgccgccg tggtgtgggc ccgcaagatc 60
cccaaggtgg gccacacctt cttccagaag cccgagagct gcccccccgt gcccggcggc 120
agcatgaagc tggacatcgg catcatcaac gagaaccagc gcgtgagcat gagccgcaac 180
atcgagagcc gcagcaccag cccctggaac tacaccgtga cctgggaccc caaccgctac 240
cccagcgagg tggtgcaggc ccagtgccgc aacctgggct gcatcaacgc ccagggcaag 300
gaggacatca gcatgaacag cgtgcccatc cagcaggaga ccctggtggt gcgccgcaag 360
caccagggct gcagcgtgag cttccagctg gagaaggtgc tggtgaccgt gggctgcacc 420
tgcgtgaccc ccgtgatcca ccacgtgcag ggcagccacc accaccacca ccactaa 477
<210> 47
<211> 1644
<212> DNA
<213> Artificial Sequence
<220>
<223> sequence encoding Signal peptide + Human CD73+ poly His tag of the present invention
<400> 47
atgagcgccc tgctgatcct ggccctggtg ggcgccgccg tggtgtgggc ctgggagctg 60
accatcctgc acaccaacga cgtgcacagc cgcctggagc agaccagcga ggacagcagc 120
aagtgcgtga acgccagccg ctgcatgggc ggcgtggccc gcctgttcac caaggtgcag 180
cagatccgcc gcgccgagcc caacgtgctg ctgctggacg ccggcgacca gtaccagggc 240
accatctggt tcaccgtgta caagggcgcc gaggtggccc acttcatgaa cgccctgcgc 300
tacgacgcca tggccctggg caaccacgag ttcgacaacg gcgtggaggg cctgatcgag 360
cccctgctga aggaggccaa gttccccatc ctgagcgcca acatcaaggc caagggcccc 420
gccagccaga tcagcggcct gtacctgccc tacaaggtgc tgcccgtggg cgacgaggtg 480
gtgggcatcg tgggctacac cagcaaggag acccccttcc tgagcaaccc cggcaccaac 540
ctggtgttcg aggacgagat caccgccctg cagcccgagg tggacaagct gaagaccctg 600
aacgtgaaca agatcatcgc cctgggccac agcggcttcg agatggacaa gctgatcgcc 660
cagaaggtgc gcggcgtgga cgtggtggtg ggcggccaca gcaacacctt cctgtacacc 720
ggcaaccccc ccagcaagga ggtgcccgcc ggcaagtacc ccttcatcgt gaccagcgac 780
gacggccgca aggtgcccgt ggtgcaggcc tacgccttcg gcaagtacct gggctacctg 840
aagatcgagt tcgacgagcg cggcaacgtg atcagcagcc acggcaaccc catcctgctg 900
aacagcagca tccccgagga ccccagcatc aaggccgaca tcaacaagtg gcgcatcaag 960
ctggacaact acagcaccca ggagctgggc aagaccatcg tgtacctgga cggcagcagc 1020
cagagctgcc gcttccgcga gtgcaacatg ggcaacctga tctgcgacgc catgatcaac 1080
aacaacctgc gccacaccga cgagatgttc tggaaccacg tgagcatgtg catcctgaac 1140
ggcggcggca tccgcagccc catcgacgag cgcaacaacg gcaccatcac ctgggagaac 1200
ctggccgccg tgctgccctt cggcggcacc ttcgacctgg tgcagctgaa gggcagcacc 1260
ctgaagaagg ccttcgagca cagcgtgcac cgctacggcc agagcaccgg cgagttcctg 1320
caggtgggcg gcatccacgt ggtgtacgac ctgagccgca agcccggcga ccgcgtggtg 1380
aagctggacg tgctgtgcac caagtgccgc gtgcccagct acgaccccct gaagatggac 1440
gaggtgtaca aggtgatcct gcccaacttc ctggccaacg gcggcgacgg cttccagatg 1500
atcaaggacg agctgctgcg ccacgacagc ggcgaccagg acatcaacgt ggtgagcacc 1560
tacatcagca agatgaaggt gatctacccc gccgtggagg gccgcatcaa gttcagcggc 1620
agccaccacc accaccacca ctaa 1644
<210> 48
<211> 1422
<212> DNA
<213> Artificial Sequence
<220>
<223> sequence encoding self signal peptide + Human IL-17RC + poly His tag
<400> 48
atgcccgtgc cctggttcct gctgagcctg gccctgggcc gcagccccgt ggtgctgagc 60
ctggagcgcc tggtgggccc ccaggacgcc acccactgca gccccggcct gagctgccgc 120
ctgtgggaca gcgacatcct gtgcctgccc ggcgacatcg tgcccgcccc cggccccgtg 180
ctggccccca cccacctgca gaccgagctg gtgctgcgct gccagaagga gaccgactgc 240
gacctgtgcc tgcgcgtggc cgtgcacctg gccgtgcacg gccactggga ggagcccgag 300
gacgaggaga agttcggcgg cgccgccgac agcggcgtgg aggagccccg caacgccagc 360
ctgcaggccc aggtggtgct gagcttccag gcctacccca ccgcccgctg cgtgctgctg 420
gaggtgcagg tgcccgccgc cctggtgcag ttcggccaga gcgtgggcag cgtggtgtac 480
gactgcttcg aggccgccct gggcagcgag gtgcgcatct ggagctacac ccagccccgc 540
tacgagaagg agctgaacca cacccagcag ctgcccgact gccgcggcct ggaggtgtgg 600
aacagcatcc ccagctgctg ggccctgccc tggctgaacg tgagcgccga cggcgacaac 660
gtgcacctgg tgctgaacgt gagcgaggag cagcacttcg gcctgagcct gtactggaac 720
caggtgcagg gcccccccaa gccccgctgg cacaagaacc tgaccggccc ccagatcatc 780
accctgaacc acaccgacct ggtgccctgc ctgtgcatcc aggtgtggcc cctggagccc 840
gacagcgtgc gcaccaacat ctgccccttc cgcgaggacc cccgcgccca ccagaacctg 900
tggcaggccg cccgcctgca gctgctgacc ctgcagagct ggctgctgga cgccccctgc 960
agcctgcccg ccgaggccgc cctgtgctgg cgcgcccccg gcggcgaccc ctgccagccc 1020
ctggtgcccc ccctgagctg ggagaacgtg accgtggaca aggtgctgga gttccccctg 1080
ctgaagggcc accccaacct gtgcgtgcag gtgaacagca gcgagaagct gcagctgcag 1140
gagtgcctgt gggccgacag cctgggcccc ctgaaggacg acgtgctgct gctggagacc 1200
cgcggccccc aggacaaccg cagcctgtgc gccctggagc ccagcggctg caccagcctg 1260
cccagcaagg ccagcacccg cgccgcccgc ctgggcgagt acctgctgca ggacctgcag 1320
agcggccagt gcctgcagct gtgggacgac gacctgggcg ccctgtgggc ctgccccatg 1380
gacaagtaca tccacggcag ccaccaccac caccaccact aa 1422
<210> 49
<211> 483
<212> DNA
<213> Artificial Sequence
<220>
<223> sequence encoding self signal peptide + Human CD28+ poly His tag
<400> 49
atgctgcgcc tgctgctggc cctgaacctg ttccccagca tccaggtgac cggcaacaag 60
atcctggtga agcagagccc catgctggtg gcctacgaca acgccgtgaa cctgagctgc 120
aagtacagct acaacctgtt cagccgcgag ttccgcgcca gcctgcacaa gggcctggac 180
agcgccgtgg aggtgtgcgt ggtgtacggc aactacagcc agcagctgca ggtgtacagc 240
aagaccggct tcaactgcga cggcaagctg ggcaacgaga gcgtgacctt ctacctgcag 300
aacctgtacg tgaaccagac cgacatctac ttctgcaaga tcgaggtgat gtaccccccc 360
ccctacctgg acaacgagaa gagcaacggc accatcatcc acgtgaaggg caagcacctg 420
tgccccagcc ccctgttccc cggccccagc aagcccggca gccaccacca ccaccaccac 480
taa 483
<210> 50
<211> 528
<212> DNA
<213> Artificial Sequence
<220>
<223> sequence encoding self signal peptide + Human PD-1+ poly His tag
<400> 50
atgcagatcc cccaggcccc ctggcccgtg gtgtgggccg tgctgcagct gggctggcgc 60
cccggctggt tcctggacag ccccgaccgc ccctggaacc cccccacctt cagccccgcc 120
ctgctggtgg tgaccgaggg cgacaacgcc accttcacct gcagcttcag caacaccagc 180
gagagcttcg tgctgaactg gtaccgcatg agccccagca accagaccga caagctggcc 240
gccttccccg aggaccgcag ccagcccggc caggactgcc gcttccgcgt gacccagctg 300
cccaacggcc gcgacttcca catgagcgtg gtgcgcgccc gccgcaacga cagcggcacc 360
tacctgtgcg gcgccatcag cctggccccc aaggcccaga tcaaggagag cctgcgcgcc 420
gagctgcgcg tgaccgagcg ccgcgccgag gtgcccaccg cccaccccag ccccagcccc 480
cgccccgccg gccagttcca gggcagccac caccaccacc accactaa 528
<210> 51
<211> 447
<212> DNA
<213> Artificial Sequence
<220>
<223> sequence encoding self signal peptide + Human ICOS + poly His tag
<400> 51
atgaagagcg gcctgtggta cttcttcctg ttctgcctgc gcatcaaggt gctgaccggc 60
gagatcaacg gcagcgccaa ctacgagatg ttcatcttcc acaacggcgg cgtgcagatc 120
ctgtgcaagt accccgacat cgtgcagcag ttcaagatgc agctgctgaa gggcggccag 180
atcctgtgcg acctgaccaa gaccaagggc agcggcaaca ccgtgagcat caagagcctg 240
aagttctgcc acagccagct gagcaacaac agcgtgagct tcttcctgta caacctggac 300
cacagccacg ccaactacta cttctgcaac ctgagcatct tcgacccccc ccccttcaag 360
gtgaccctga ccggcggcta cctgcacatc tacgagagcc agctgtgctg ccagctgaag 420
ggcagccacc accaccacca ccactaa 447
<210> 52
<211> 1104
<212> DNA
<213> Artificial Sequence
<220>
<223> sequence encoding self signal peptide + Human Decorin + poly His tag
<400> 52
atgaaggcca ccatcatcct gctgctgctg gcccaggtga gctgggccgg ccccttccag 60
cagcgcggcc tgttcgactt catgctggag gacgaggcca gcggcatcgg ccccgaggtg 120
cccgacgacc gcgacttcga gcccagcctg ggccccgtgt gccccttccg ctgccagtgc 180
cacctgcgcg tggtgcagtg cagcgacctg ggcctggaca aggtgcccaa ggacctgccc 240
cccgacacca ccctgctgga cctgcagaac aacaagatca ccgagatcaa ggacggcgac 300
ttcaagaacc tgaagaacct gcacgccctg atcctggtga acaacaagat cagcaaggtg 360
agccccggcg ccttcacccc cctggtgaag ctggagcgcc tgtacctgag caagaaccag 420
ctgaaggagc tgcccgagaa gatgcccaag accctgcagg agctgcgcgc ccacgagaac 480
gagatcacca aggtgcgcaa ggtgaccttc aacggcctga accagatgat cgtgatcgag 540
ctgggcacca accccctgaa gagcagcggc atcgagaacg gcgccttcca gggcatgaag 600
aagctgagct acatccgcat cgccgacacc aacatcacca gcatccccca gggcctgccc 660
cccagcctga ccgagctgca cctggacggc aacaagatca gccgcgtgga cgccgccagc 720
ctgaagggcc tgaacaacct ggccaagctg ggcctgagct tcaacagcat cagcgccgtg 780
gacaacggca gcctggccaa caccccccac ctgcgcgagc tgcacctgga caacaacaag 840
ctgacccgcg tgcccggcgg cctggccgag cacaagtaca tccaggtggt gtacctgcac 900
aacaacaaca tcagcgtggt gggcagcagc gacttctgcc cccccggcca caacaccaag 960
aaggccagct acagcggcgt gagcctgttc agcaaccccg tgcagtactg ggagatccag 1020
cccagcacct tccgctgcgt gtacgtgcgc agcgccatcc agctgggcaa ctacaagggc 1080
agccaccacc accaccacca ctaa 1104
<210> 53
<211> 810
<212> DNA
<213> Artificial Sequence
<220>
<223> sequence encoding self signal peptide + Human Siglec-15+ poly His tag
<400> 53
atggagaaga gcatctggct gctggcctgc ctggcctggg tgctgcccac cggcagcttc 60
gtgcgcacca agatcgacac caccgagaac ctgctgaaca ccgaggtgca cagcagcccc 120
gcccagcgct ggagcatgca ggtgcccccc gaggtgagcg ccgaggccgg cgacgccgcc 180
gtgctgccct gcaccttcac ccacccccac cgccactacg acggccccct gaccgccatc 240
tggcgcgccg gcgagcccta cgccggcccc caggtgttcc gctgcgccgc cgcccgcggc 300
agcgagctgt gccagaccgc cctgagccac ggccgcttcc gcctgctggg caacccccgc 360
cgcaacgacc tgagcctgcg cgtggagcgc ctggccctgg ccgacgaccg ccgctacttc 420
tgccgcgtgg agttcgccgg cgacgtgcac gaccgctacg agagccgcca cggcgtgcgc 480
ctgcacgtga ccgccgcccc ccgcatcgtg aacatcagcg tgctgcccag ccccgcccac 540
gccttccgcg ccctgtgcac cgccgagggc gagccccccc ccgccctggc ctggagcggc 600
cccgccctgg gcaacagcct ggccgccgtg cgcagccccc gcgagggcca cggccacctg 660
gtgaccgccg agctgcccgc cctgacccac gacggccgct acacctgcac cgccgccaac 720
agcctgcgca gcgaggccag cgtgtacctg ttccgcttcc acggcgccag cggcgccagc 780
accggcagcc accaccacca ccaccactaa 810
<210> 54
<211> 558
<212> DNA
<213> Artificial Sequence
<220>
<223> sequence encoding self signal peptide + Human IL-7+ poly His tag
<400> 54
atgttccacg tgagcttccg ctacatcttc ggcctgcccc ccctgatcct ggtgctgctg 60
cccgtggcca gcagcgactg cgacatcgag ggcaaggacg gcaagcagta cgagagcgtg 120
ctgatggtga gcatcgacca gctgctggac agcatgaagg agatcggcag caactgcctg 180
aacaacgagt tcaacttctt caagcgccac atctgcgacg ccaacaagga gggcatgttc 240
ctgttccgcg ccgcccgcaa gctgcgccag ttcctgaaga tgaacagcac cggcgacttc 300
gacctgcacc tgctgaaggt gagcgagggc accaccatcc tgctgaactg caccggccag 360
gtgaagggcc gcaagcccgc cgccctgggc gaggcccagc ccaccaagag cctggaggag 420
aacaagagcc tgaaggagca gaagaagctg aacgacctgt gcttcctgaa gcgcctgctg 480
caggagatca agacctgctg gaacaagatc ctgatgggca ccaaggagca cggcagccac 540
caccaccacc accactaa 558
<210> 55
<211> 516
<212> DNA
<213> Artificial Sequence
<220>
<223> sequence encoding self signal peptide + HumanIL-17F + poly His tag
<400> 55
atgaccgtga agaccctgca cggccccgcc atggtgaagt acctgctgct gagcatcctg 60
ggcctggcct tcctgagcga ggccgccgcc cgcaagatcc ccaaggtggg ccacaccttc 120
ttccagaagc ccgagagctg cccccccgtg cccggcggca gcatgaagct ggacatcggc 180
atcatcaacg agaaccagcg cgtgagcatg agccgcaaca tcgagagccg cagcaccagc 240
ccctggaact acaccgtgac ctgggacccc aaccgctacc ccagcgaggt ggtgcaggcc 300
cagtgccgca acctgggctg catcaacgcc cagggcaagg aggacatcag catgaacagc 360
gtgcccatcc agcaggagac cctggtggtg cgccgcaagc accagggctg cagcgtgagc 420
ttccagctgg agaaggtgct ggtgaccgtg ggctgcacct gcgtgacccc cgtgatccac 480
cacgtgcagg gcagccacca ccaccaccac cactaa 516
<210> 56
<211> 1671
<212> DNA
<213> Artificial Sequence
<220>
<223> sequence encoding self signal peptide + Human CD73+ poly His tag
<400> 56
atgtgccccc gcgccgcccg cgcccccgcc accctgctgc tggccctggg cgccgtgctg 60
tggcccgccg ccggcgcctg ggagctgacc atcctgcaca ccaacgacgt gcacagccgc 120
ctggagcaga ccagcgagga cagcagcaag tgcgtgaacg ccagccgctg catgggcggc 180
gtggcccgcc tgttcaccaa ggtgcagcag atccgccgcg ccgagcccaa cgtgctgctg 240
ctggacgccg gcgaccagta ccagggcacc atctggttca ccgtgtacaa gggcgccgag 300
gtggcccact tcatgaacgc cctgcgctac gacgccatgg ccctgggcaa ccacgagttc 360
gacaacggcg tggagggcct gatcgagccc ctgctgaagg aggccaagtt ccccatcctg 420
agcgccaaca tcaaggccaa gggccccgcc agccagatca gcggcctgta cctgccctac 480
aaggtgctgc ccgtgggcga cgaggtggtg ggcatcgtgg gctacaccag caaggagacc 540
cccttcctga gcaaccccgg caccaacctg gtgttcgagg acgagatcac cgccctgcag 600
cccgaggtgg acaagctgaa gaccctgaac gtgaacaaga tcatcgccct gggccacagc 660
ggcttcgaga tggacaagct gatcgcccag aaggtgcgcg gcgtggacgt ggtggtgggc 720
ggccacagca acaccttcct gtacaccggc aaccccccca gcaaggaggt gcccgccggc 780
aagtacccct tcatcgtgac cagcgacgac ggccgcaagg tgcccgtggt gcaggcctac 840
gccttcggca agtacctggg ctacctgaag atcgagttcg acgagcgcgg caacgtgatc 900
agcagccacg gcaaccccat cctgctgaac agcagcatcc ccgaggaccc cagcatcaag 960
gccgacatca acaagtggcg catcaagctg gacaactaca gcacccagga gctgggcaag 1020
accatcgtgt acctggacgg cagcagccag agctgccgct tccgcgagtg caacatgggc 1080
aacctgatct gcgacgccat gatcaacaac aacctgcgcc acaccgacga gatgttctgg 1140
aaccacgtga gcatgtgcat cctgaacggc ggcggcatcc gcagccccat cgacgagcgc 1200
aacaacggca ccatcacctg ggagaacctg gccgccgtgc tgcccttcgg cggcaccttc 1260
gacctggtgc agctgaaggg cagcaccctg aagaaggcct tcgagcacag cgtgcaccgc 1320
tacggccaga gcaccggcga gttcctgcag gtgggcggca tccacgtggt gtacgacctg 1380
agccgcaagc ccggcgaccg cgtggtgaag ctggacgtgc tgtgcaccaa gtgccgcgtg 1440
cccagctacg accccctgaa gatggacgag gtgtacaagg tgatcctgcc caacttcctg 1500
gccaacggcg gcgacggctt ccagatgatc aaggacgagc tgctgcgcca cgacagcggc 1560
gaccaggaca tcaacgtggt gagcacctac atcagcaaga tgaaggtgat ctaccccgcc 1620
gtggagggcc gcatcaagtt cagcggcagc caccaccacc accaccacta a 1671
Claims (10)
1. A polypeptide of (a) or (b) below:
(a) polypeptide consisting of an amino acid sequence shown in SEQ ID No. 1;
(b) the polypeptide which is formed by substituting, deleting and/or adding one or more amino acids in the amino acid sequence shown in SEQ ID No.1 and has the same function with the polypeptide formed by the amino acid sequence shown in SEQ ID No. 1.
2. The polypeptide of claim 1, wherein the polypeptide consisting of the amino acid sequence shown in SEQ ID No.1 with one or more amino acid substitutions, deletions and/or additions and having the same function as the polypeptide shown in SEQ ID No.1 at least comprises a sequence of at least 9 consecutive amino acids, preferably at least 10 amino acids, more preferably at least 11 amino acids at the C-terminal end of the amino acid sequence shown in SEQ ID No. 1.
3. A polynucleotide encoding the polypeptide of claim 1 or 2.
4. An expression vector comprising the polynucleotide of claim 3.
5. A mammalian cell comprising the polynucleotide of claim 3 or the expression vector of claim 4.
6. Use of the polypeptide of claim 1 or 2 as a signal peptide for secretory expression of a protein of interest in a mammalian cell.
7. A method for secretory expression of a protein of interest in a mammalian cell, the method comprising:
secretory expression of a target protein in a mammalian cell containing the polynucleotide of claim 3 or the expression vector of claim 4.
8. The method of claim 7, wherein the mammalian cells are selected from one or more of Chinese Hamster Ovary (CHO), mouse plasmacytoma (A non-Ig differentiation, non-light-synthesizing subclone of NS-1, NSO), Baby Hamster Kidney (BHK), human embryonic kidney293 (HEK 293) cells.
9. The method according to claim 7, wherein the protein of interest is selected from one or more of Human IL-17RC, Human CD28, Human PD-1, Human ICOS, Human Decorin, Human Siglec-15, Human IL-7, Human IL-17F, and Human CD 73.
10. The method of claim 7, comprising:
ligating a polynucleotide encoding the polypeptide of claim 1 or 2 with a polynucleotide encoding a protein of interest, said polynucleotide encoding the polypeptide of claim 1 or 2 being positioned at the front end of said polynucleotide encoding the protein of interest, and cloning into a mammalian cell expression vector;
and (3) transfecting the expression vector into a mammalian cell to express the target protein.
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Cited By (2)
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CN113817709A (en) * | 2021-08-12 | 2021-12-21 | 中国科学院天津工业生物技术研究所 | Carbohydrate-binding domain CBM68 and uses thereof |
CN113925876A (en) * | 2021-10-14 | 2022-01-14 | 北京创世客生物技术有限公司 | Use of CIK immune cells in the treatment of cancer |
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CN104968795A (en) * | 2012-12-05 | 2015-10-07 | 斯特拉塔吉亚医疗公司 | Protein expression enhancing polypeptides |
CN104974226A (en) * | 2014-04-01 | 2015-10-14 | 上海中信国健药业股份有限公司 | Signal peptide for protein expression |
CN106478773A (en) * | 2015-08-25 | 2017-03-08 | 三生国健药业(上海)股份有限公司 | A kind of novel signal peptide of synthetic |
CN110452288A (en) * | 2018-05-08 | 2019-11-15 | 鲁南制药集团股份有限公司 | A kind of artificial synthesized signal peptide |
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CN104968795A (en) * | 2012-12-05 | 2015-10-07 | 斯特拉塔吉亚医疗公司 | Protein expression enhancing polypeptides |
CN104974226A (en) * | 2014-04-01 | 2015-10-14 | 上海中信国健药业股份有限公司 | Signal peptide for protein expression |
CN106478773A (en) * | 2015-08-25 | 2017-03-08 | 三生国健药业(上海)股份有限公司 | A kind of novel signal peptide of synthetic |
CN110452288A (en) * | 2018-05-08 | 2019-11-15 | 鲁南制药集团股份有限公司 | A kind of artificial synthesized signal peptide |
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CN113817709A (en) * | 2021-08-12 | 2021-12-21 | 中国科学院天津工业生物技术研究所 | Carbohydrate-binding domain CBM68 and uses thereof |
CN113817709B (en) * | 2021-08-12 | 2023-08-04 | 中国科学院天津工业生物技术研究所 | Carbohydrate binding domain CBM68 and uses thereof |
CN113925876A (en) * | 2021-10-14 | 2022-01-14 | 北京创世客生物技术有限公司 | Use of CIK immune cells in the treatment of cancer |
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