CN1532284A - Novel antibiotic polypeptide and its preparing method - Google Patents

Novel antibiotic polypeptide and its preparing method Download PDF

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CN1532284A
CN1532284A CNA031204007A CN03120400A CN1532284A CN 1532284 A CN1532284 A CN 1532284A CN A031204007 A CNA031204007 A CN A031204007A CN 03120400 A CN03120400 A CN 03120400A CN 1532284 A CN1532284 A CN 1532284A
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丘小庆
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YANGHUI BIOLOGICAL SCIENCE AND TECHNOLOGY Co Ltd CHENGDU CITY
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YANGHUI BIOLOGICAL SCIENCE AND TECHNOLOGY Co Ltd CHENGDU CITY
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Priority claimed from CNA021340420A external-priority patent/CN1513877A/en
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Priority to PCT/CN2004/000046 priority patent/WO2004082701A1/en
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Abstract

The present invention discloses a kind of novel antibiotic polypeptide and its preparation process. The recombinant antibiotic polypeptide contains colicine capable of forming ion channel or its aquatic porous channel structure domain and staphylococcal bacteria signal transducing polypeptide. The present invention also discloses the nucleotide sequence encoding the recombinant antibiotic polypeptide and recombinant plasmid containing said nucleotide sequence. Compared with traditional antibiotics, the novel antibiotic polypeptide of the present invention has the advantage of no drug resistance the bacteria are induced to produce. The bacteria are hard to alter their phosphatide double molecular structure via mutation, and the recombinant polypeptide of the present invention reaches the aim of killing bacteria via forming ion channel on the cell membrane of target bacteria directly.

Description

A kind of novel antibacterial polypeptide and preparation method thereof
Technical field
The present invention relates to a kind of staphylococcus polypeptide of reorganization, the Nucleotide of this polypeptide of encoding and aminoacid sequence.
The invention still further relates to the preparation method of reorganization staphylococcus polypeptide.
Background technology
Infectation of bacteria is human life and healthy main threat, since sulfanilamide (SN) and penicillin come out, the antibiotic of human invention successively mainly with by suppress bacterial cell wall synthetic, suppress or disturb the nucleic acid of bacterium and proteinic metabolism and route of synthesis to reach antibiotic purpose.Yet these antibiotic approach lure that easily bacterium undergos mutation and produce antibiotic resistance into.Therefore people are being devoted to the antibiotic of development of new always.Directly forming ionic channel and cause bacterium death on bacterial membrane is one of more promising antibiotic exploitation direction.Occurring in nature, bacteriotoxin quite a lot comes killing bacteria with this mechanism, its type specimen is exactly a kind of toxin protein of intestinal bacteria (Escherichia coli) excretory a---colicin (colicin), nineteen fifty-two Jacob finds some bacillus that its intestinal bacteria that can kill other strain system specifically and relevant strain are, as Shigella Sonnei (Jacob et al,) Sur labiosynthese d ' une colicine et son mode d ' action, Annals of thePasteur Institute.83:295-315 (1952)).Finkelstein in 1978 etc. have found that the colicin that can form ionic channel can form the voltage-dependent ionic channel on artificial bimolecular lipid membrane, thereby at the antibacterial mechanisms that has fundamentally disclosed this bacterioid toxin (Schein et al .Colicin K acts by forming voltagedependent channels in phospholipid bilayer membranes.Nature 276:159-163 (1973)).Qiu in 1996 and Finkelstein etc. have disclosed ionic channel that colicin Ia forms on artificial bimolecular lipid membrane open and stride film three-dimensional arrangement (Qiu et al. when closing, Major transmembrane movementassociated with colicin, Ia channel gating.J.Gen.Physiology.107:313-328 (1996)), established theoretical basis on molecular level, designing and prepare novel antibiotic.
Recent two decades, people find that gradually bacterium also has the excretory signal conduction polypeptide with the external environment variation, regulating proteic gene in the after birth acceptor of preparation signal conduction polypeptide and the born of the same parents often is coupled at together, controlling growth and the division of bacterium in phase, but the research of this direction is made slow progress always, signal conduction polypeptide A grD such as staphylococcus aureus, until nineteen ninety-five is just made clear at last is an octapeptide (Ji et al., Celldensity control of staphylococcal virulence mediated by an octapeptide pheromone.Proc.Natl.Acad.Sci.USA 92:12055-12059 (1995)).Found that this octapeptide of the early stage of streptococcus aureus logarithmic phase or middle and advanced stage adding synthetic can stimulate or suppress growth (the Mayville et al. of streptococcus aureus in 1999, Structure activity analysis of synthetic autoinducingthiolactone peptides from staphylococcus aureus resoonsible for virulence.PNAS, 96:1218-1223 (1999)).
As mentioned above, colicin is a kind of ideal ionic channel antibiotic, shortcoming is to act on Gram-negative bacillus such as intestinal bacteria, if can utilize the distinctive signaling peptides of pathogenic bacterium as inductor, with colicin or its water-based pore passage structure or be induced to and form ionic channel near the specific bacterial membrane and kill and wound this pathogenic bacterium, should be a kind of ideal antibiotic exploitation direction.
Summary of the invention
One object of the present invention is to provide a kind of novel reorganization antimicrobial polypeptide, and it can avoid bringing out bacterium
Another object of the present invention is to provide the nucleotide sequence of this reorganization antimicrobial polypeptide of coding.
A further object of the present invention is to provide a recombinant plasmid, and it contains the nucleotide sequence of code book invention reorganization antimicrobial polypeptide.
A further object of the present invention is to provide the monoamino-acid sequence, its reorganization antimicrobial polypeptide of the present invention of encoding.
A further object of the present invention is to provide and contains the recombinate pharmaceutical composition of antimicrobial polypeptide of the present invention.
A further object of the present invention is to provide the preparation method of reorganization antimicrobial polypeptide.
According to an aspect of the present invention, the gene with coding staphylococcus signal conduction polypeptide is connected the gene of staphylococcus polypeptide thereby the acquisition coding is recombinated with the colicin gene.
In a preferred embodiment of the invention, adopt the signal of coding streptococcus aureus and staphylococcus epidermidis to conduct the pheromone gene of the gene of polypeptide as antimicrobial polypeptide respectively, the gene of the golden yellow grape signal conduction polypeptide of wherein encoding is selected from Agr D I (SEQ ID NO.1), Agr D II (SEQ ID NO.3) and Agr DIII (SEQ ID NO.5), Agr D IV (SEQ ID NO.7); The gene of coding staphylococcus epidermidis signal conduction polypeptide is the gene with SEQ ID NO.9 nucleotide sequence.The optional self energy of colicin forms colicin El, Ia, Ib, A, B and N or its water-based pore passage structure territory of ionic channel, in a preferred embodiment of the invention, above-mentioned signal conduction polypeptide gene is connected with the carboxyl terminal of colicin Ia (SEQ ID NO.11) or aminoterminal respectively and forms nucleotide sequence as SEQ ID NO.12, SEQ ID NO.14, SEQ ID NO.16, SEQ ID NO.18, SEQ IDNO.20 and SEQ ID NO.22.These nucleotide sequence coded aminoacid sequences as SEQ ID NO.13, SEQ ID NO.15, SEQ ID NO.17, SEQ ID NO.19, SEQID NO.21 and SEQ ID NO.23, these aminoacid sequence encoded polypeptide promptly correspond to antimicrobial polypeptide Ph-SA1 of the present invention respectively, Ph-SA2, Ph-SA3, Ph-SA4, Ph-SE, and Ph-SAR1.
In the reorganization antimicrobial polypeptide that makes up, can induce colicin channel architecture territory to pass near bacterial outer membrane and the intermembrane space arrival target cell membrane as inductor with the signal of target bacteria after birth receptors bind conduction polypeptide, water-based pore passage structure territory in the colicin channel architecture territory forms ionic channel on the target bacteria after birth then, make content leaks in the target bacteria born of the same parents and cause target bacteria death, thereby reach germ-resistant purpose.
The expressed aminoacid sequence of the coded aminoacid sequence of nucleotide sequence SEQ ID NO.1 (SEQ ID NO:2) of coding streptococcus aureus signal conduction polypeptide and Genebank U85097 sequence is identical among the present invention; The expressed aminoacid sequence of the coded aminoacid sequence of nucleotide sequence SEQ ID NO.3 (SEQ ID NO:4) of coding streptococcus aureus signal conduction polypeptide and Genebank AF001782 sequence is identical; The expressed aminoacid sequence of the coded aminoacid sequence of nucleotide sequence SEQ ID NO.5 (SEQ ID NO:6) of coding streptococcus aureus signal conduction polypeptide and Genebank AF001783 sequence is identical; The expressed aminoacid sequence of the coded aminoacid sequence of nucleotide sequence SEQ ID NO.7 (SEQ ID NO:8) of coding streptococcus aureus signal conduction polypeptide and Genebank AF288215 sequence is identical; The expressed aminoacid sequence of the coded aminoacid sequence of nucleotide sequence SEQ ID NO.9 (SEQ ID NO:10) of coding staphylococcus epidermidis signal conduction polypeptide and Genebank Z49220 sequence is identical.
According to a further aspect in the invention, provide as Fig. 1～plasmid vector that comprises Nucleotide of the present invention shown in Figure 6, this plasmid vector is the nucleotide sequence of aforesaid coding staphylococcus signal conduction polypeptide to be inserted through double-stranded oligonucleotide point mutation technology form recombinant plasmid of the present invention on the 626th amino acids of colicin Ia gene or the 1st amino acids.
Among the present invention, the original plasmid pSELECTTM-1 that makes up plasmid vector comes from Promega company, colicin Ia and immunity protein gene have wherein been loaded, at streptococcus aureus signal conduction gene design four pairs of primers, its sequence is respectively shown in SEQ ID NO.:24～SEQ ID NO:32; At epidermis grape ball signal conduction gene design a pair of primer, its sequence is shown in SEQ ID NO:33 and SEQ IDNO:34.Utilize double-stranded oligonucleotide point mutation technology, obtain recombinant plasmid, again the recombinant plasmid that obtains is transfected into the e. coli tg1 engineering bacteria and obtains to produce the engineering bacteria cell of reorganization antimicrobial polypeptide according to the operation of Strategene company medicine-chest.
In accordance with a further aspect of the present invention, provide and contain the recombinate pharmaceutical composition of antimicrobial polypeptide of the present invention, can make the pharmaceutical composition that is suitable for clinical use by polypeptide of the present invention is added pharmaceutically acceptable carrier or vehicle or optional additional components.
According to another aspect of the invention, the preparation method of antimicrobial polypeptide of the present invention is provided, the gene of coding staphylococcus signal conduction polypeptide operationally is connected the gene that obtains coding reorganization antimicrobial polypeptide with the colicin gene, the gene that obtains imported in the expression system express, separate polypeptide expressed and obtain antimicrobial polypeptide of the present invention.
Antimicrobial polypeptide of the present invention is that compared to the advantage of traditional antibiotic it can not induce bacterium to produce traditional resistance.As everyone knows, bacterium can produce β-Nei Xiananmei by sudden change, reduce modes such as taking in, change the drug effect site change its cell wall structure, and the metabolism etc. that changes its protein and nucleic acid produces resistance to traditional antibiotic.But bacterium but is difficult to change by sudden change the structure of the phospholipid bilayer of its cytolemma, and this is distinctive feature of the present invention just, and recombinant polypeptide of the present invention reaches the sterilization purpose by directly form ionic channel on the after birth of target bacteria.
Brief description of drawings
Fig. 1 illustrates the structure of the plasmid pCHCSA1 that contains streptococcus aureus signal conduction polypeptide A gr D I and colicin Ia;
Fig. 2 illustrates the structure of the plasmid pCHCSA2 that contains streptococcus aureus signal conduction polypeptide A gr D II and colicin Ia;
Fig. 3 illustrates the structure of the plasmid pCHCSA3 that contains streptococcus aureus signal conduction polypeptide A gr D III and colicin Ia;
Fig. 4 illustrates the structure of the plasmid pCHCSA4 that contains streptococcus aureus signal conduction polypeptide A gr D IV and colicin Ia;
Fig. 5 illustrates the structure of the plasmid pCHCSE that contains staphylococcus epidermidis signal conduction polypeptide and colicin Ia;
Fig. 6 illustrates the plasmid structure that contains streptococcus aureus signal conduction polypeptide A grD I and colicin Ia, and wherein Agr D I is connected on the 1st amino acids of colicin Ia;
Fig. 7 illustrates the structure of the antimicrobial polypeptide that comprises 634 amino-acid residues;
Fig. 8 is the inhibition experimental result of antimicrobial polypeptide of the present invention (Ph-SA1) to the responsive golden Portugal of penicillin bacterium, shows that the growth of the responsive golden Portugal of penicillin bacterium can only be suppressed by penicillin and Ph-SA1.Control among the figure, contrast, PEN, penicillin, COL, the wild-type colicin, TG1, the TG1 engineering bacteria albumen of plasmid-free, FusionProtein, colicin Ia carboxyl terminal has connected the recombinant protein of other octapeptide, and PMC is Ph-SA, anti-golden Portugal bacterium polypeptide.The drug dose of each experimental group is 100ng/ml;
Fig. 9 is the transmission electron microscope observing (amplify 25,000 times) of anti-golden Portugal's bacterium polypeptide (Ph-SA1) to methicillin-resistant staphylococcus aureus (ATCC BAA-42) sterilization effect.Control, control group, normal morphology bacterium; Wildtype colicin Ia, wild-type colicin Ia handles (50 μ g/ml), handles after two hours, and the bacterium form is still normal; AgrD, golden Portugal bacterium signal polypeptide is handled (10 μ g/ml), handles after two hours, and ne ar is still normal, and the thalline in upper end divides; Oxacillin, Oxazacillin is handled (500 μ g/ml), and heavy dose of Oxazacillin was handled after 2 hours, and the mycetocyte wall is the dissolving form; Ph-SA1, anti-golden Portugal polypeptide is handled (50 μ g/ml), and after anti-golden Portugal polypeptide was handled half an hour, bacterium was broken, content leaks, and cell wall structures is complete;
Conclusion: use signal conduction polypeptide or wild-type colicin to handle merely and all can not kill MRSA, it is different fully with Oxazacillin that anti-golden Portugal polypeptide but can be killed MRSA and bactericidal mechanism;
Figure 10 is the survival curve of anti-golden Portugal polypeptide to MRSA (BAA42) infecting mouse endogenous protective test-results;
Figure 11 be antimicrobial polypeptide of the present invention (Ph-SA1 and Ph-SE) to the staphylococcic restraining effect of epidermis, show that the growth of staphylococcus epidermidis can be suppressed by anti-golden Portugal polypeptide portion, can effectively be suppressed by anti-table Portugal polypeptide.Among the figure: Control, contrast, PEN, penicillin, Ph-SA1, anti-golden Portugal polypeptide, Ph-SE, anti-table Portugal polypeptide.Drug level 2 μ g/ml.
Figure 12 is the growth-inhibiting effects of various antimicrobial polypeptides to methicillin-resistant gold Portugal bacterium, the growth that shows methicillin-resistant gold Portugal bacterium almost can not be suppressed by Oxazacillin, the fungistatic effect of Ph-SAR1 is the poorest, that the inhibition effect is best is Ph-SA1 and Ph-SA4, next is Ph-SE, and Ph-SA3 and Ph-SA2 take second place again; Control among the figure, contrast, Oxacillin, Oxazacillin, Ph-SA1, Ph-SA2, Ph-SA3, Ph-SA4 has connected the anti-golden Portugal polypeptide of four kinds of AgrD, Ph-SE respectively, anti-table Portugal polypeptide, Ph-SAR1, the AgrD1 pheromone is connected the N-terminal anti-golden Portugal of colicin Ia polypeptide, drug level 5 μ g/ml.
The embodiment of invention
Below in conjunction with accompanying drawing, describe the present invention in detail by description to preferred embodiment of the present invention.
[embodiment 1] expresses the structure and the preparation of reorganization antimicrobial polypeptide of the plasmid of antimicrobial polypeptide
Original plasmid is the pSELECT that has loaded colicin Ia and immunity protein gene TM-1 commercial plasmid (plasmid size 8.3kb, Promega company) (UCSF, P.Gosh gifts).Through double-stranded oligonucleotide point mutation technology (QuickChange TMKit, Strategeue company) the staphylococcus signal polypeptide gene of will encoding is inserted on the I626 site of colicin Ia gene, the a series of mutant plasmids that prepared antistaphylococcus engineering multipeptide, pCHCSA1, pCHCSA2, pCHCSA3, pCHCSA4 and pCHCSE and Agr DI are connected plasmid on the 1st amino acids of colicin Ia (as Fig. 1～shown in Figure 7).Mutant plasmid is transfected into E.coli TG1 engineering bacteria (AECOM, K.Jakes gifts) lining and prepares polypeptide.
The sudden change program is undertaken by Strategene QuikChange Site-Directed Mutagenesis Kit (Catalog#200518) medicine-chest handbook:
1. prepare the point mutation reactant:
5μl?10X?buffer
2 μ l (10ng) wild-type colicin plasmids
1.25 5 '-3 ' oligonucleotide primer (seeing listed primer sequence) of μ l (125ng) design
1.25 3 '-5 ' oligonucleotide primer (seeing listed primer sequence) of μ l (125ng) design
1μl?dNTP
Distilled water 50 μ l
1μl?pfu
(except that plasmid, primer and distilled water, being reagent that medicine-chest is equipped with)
2. carry out pcr amplification, amplification condition: 95 ℃ of sex change, 35 seconds, anneal 53 ℃, 70 seconds, extend 68 ℃, totally 20 circulations in 17 fens;
3. add (37 ℃, 1 hour) behind the Dpn1 restriction endonuclease 1 μ l digestion mother body D NA chain, get 1 μ l reactant and XL1-Blue competent cell 50 μ l ice and incubated 30 minutes, 42 ℃ of capable thermal shockings 45 seconds, were inserted in the ice 2 minutes again;
4. add NZY and train 220rpm behind the basic 0.5ml, 37 ℃ were shaken bacterium after 1 hour, got 50-100 μ l reactant bed board (LB training base adds 1% agar and adds 50 μ g/ml penbritins, and 37C spends the night);
5.18 choose bacterium after hour, follow Qiagene, companies such as Gibco various commercial extract the plasmid medicine-chests extract plasmids all can, order-checking is determined to suddenly change successfully.
6. the TG1 engineering bacteria of plasmid 50ng and preparation being experienced polypeptide cell 50 μ l ice incubated 30 fens, 42 ℃, thermal shocking in 90 seconds, get 50-100 μ l reactant adding LB and train basic 0.5ml, 220rpm, 37 ℃ are shaken bacterium bed board after 1 hour (LB training base adds 1% agar and adds 50 μ g/ml penbritins) and hatch picking colony after 18 hours for 37 ℃.
7. increase bacterium in a large number, 8-16 rises FB training base, 250rpm, 37 ℃, 6-8 hour; The centrifugation thalline, 4 ℃, 6000g, 20 minutes, get 4 ℃, 50mM borate buffer (2mM EDTA+2mM DTT) 50-80ml suspension thalline, add ultrasonication behind 0.2M PMSF 250 microlitres (4 ℃, 400W, 2 minutes), the broken thalline (4 ℃, 75000g, 1.5 hours) of high speed centrifugation, get supernatant and add the Vetstrep 5,000,000 deposit D NA of unit, get supernatant behind the high speed centrifugation (4 ℃, 30000g, 10 minutes) and pack the dialysis tubing of molecular weight 15000 in 4 ℃, after 4 liters of dialysed overnight of 50mM borate buffer, high speed centrifugation (4 ℃, 30000g, 10 minutes) back supernatant is splined on the CM ion exchange column, 4 ℃, 0.3M NaCl+50mM borate buffer wash-out can obtain the antimicrobial polypeptide of recombinating.Corresponding to above 6 kinds of plasmids, Ph-SA1, Ph-SA2, Ph-SA3, Ph-SA4, Ph-SE and six kinds of reorganization of Ph-SAR1 antimicrobial polypeptide have been obtained.
Designed oligonucleotide sequence in the above-mentioned preparation plasmid
pCHCSA1
5’-3’(SEQ?ID?NO：24)
gcg?aat?aag?ttc?tgg?ggt?att?TAT?TCC?ACC?TGT?GAT?TTT?ATA?ATG?taa?ata?aaa?tat
aag?aca?ggc
3’-5’(SEQ?ID?NO：25)
gcc?tgt?ctt?ata?ttt?tat?tta?CAT?TAT?AAA?ATC?ACA?GGT?GGA?ATA?aat?acc?cca?gaa?ctt
att?cgc
pCHCSA2
5’-3’(SEQ?ID?NO：26)
gcg?aat?aag?ttc?tgg?ggt?att?GGA?GTT?AAC?GCA?TGT?TCT?TCC?CTG?TTT?taa?ata
aaa?tat?aag?aca?ggc
3’-5’(SEQ?ID?NO：27)
gcc?tgt?ctt?ata?ttt?tat?tta?AAA?CAG?GGA?AGA?ACA?TGC?GTT?AAC?TCC?aat?acc?cca
gaa?ctt?att?cgc
pCHCSA3
5’-3’(SEQ?ID?NO：28)
gcg?aat?aag?ttc?tgg?ggt?att?TAT?ATA?AAC?TGT?GAT?TTT?CTTCTG?taa?ata?aaa?tat
aag?aca?ggc
3’-5’(SEQ?ID?NO：29)
gcc?tgt?ctt?ata?ttt?tat?tta?CAG?AAG?AAA?ATC?ACA?GTT?TAT?ATA?aat?acc?cca?gaa?ctt
att?cgc
pCHCSA4
5’-3’(SEQ?ID?NO：30)
gcg?aat?aag?ttc?tgg?ggt?att?TAT?TCC?ACC?TGT?TAC?TTT?ATA?ATG?taa?ata?aaa?tat
aag?aca?ggc
3’-5’(SEQ?ID?NO：31)
gcc?tgt?ctt?ata?ttt?tat?tta?CAT?TAT?AAA?GTA?ACA?GGT?GGA?ATA?aat?acc?cca?gaa?ctt
att?cgc
pCHCSE
5’-3’(SEQ?ID?NO：32)
gcg?aat?aag?ttc?tgg?ggt?att?GAT?TCC?GTT?TGT?GCA?TCC?TAT?TTT?taa?ata?aaa?tat
aag?aca?ggc
3’-5’(SEQ?ID?NO：33)
gcc?tgt?ctt?ata?ttt?tat?tta?AAA?ATA?GGA?TGC?ACA?AAC?GGA?ATC?aat?acc?cca?gaa
ctt?att?cgc
[embodiment 2] reorganization antimicrobial polypeptide is to the restraining effect of the responsive golden Portugal of penicillin bacterium
Bacterium is the USS bacterial strain, the responsive streptococcus aureus of ATCC 25923 penicillin, bacterium liquid 5 microlitres (10 8CFU/ml level bacterium amount) adds 1% Tryptones, 1%NaCl, 0.5% yeast, 0.5% glucose, 1%HK 2PO 4Nutrient solution 10ml in, prepare 6 groups altogether, first group adds 0.3M NaCl+50mM borate buffer (the antimicrobial polypeptide amount of liquid that adds in amount and the experimental group is identical) in contrast, second group adds the 100ng/ml Benzylpenicillin sodium, the 3rd group adds 100ng/ml wild-type colicin Ia (it is 0.3M NaCl+50mM borate buffer that colicin is preserved liquid), the 4th group adds the TG1 engineering bacteria albumen (preserving liquid is 0.3M NaCl+50mM borate buffer) that does not contain plasmid, the 5th group adds the fusion rotein (preserving liquid is 0.3M NaCl+50mM borate buffer) that 100ng/ml colicin Ia carboxyl terminal has connected other octapeptide, and the 6th group adds the anti-golden bacterium polypeptide Ph-SA1 of Portugal of 100ng/ml (preserving liquid is 0.3M NaCl+50mM borate buffer).
The above-mentioned liquid of respectively organizing places the 100ml Erlenmeyer flask, 200rpm, 37 ℃ of growths, the 100 μ l that per hour sample add in the 96 hole enzyme plates through spectrophotometer (A 595nm) colorimetric bacteria tested growth turbidity, the growth curve of bacteria that draws comes the inhibitory effect of comparison antimicrobial polypeptide, the result shows that the growth of the responsive golden Portugal of penicillin bacterium can only be suppressed by penicillin and Ph-SA1 as shown in Figure 8.
[embodiment 3] anti-golden Portugal bacterium polypeptide is to the transmission electron microscope observing (dyeing of 1% phospho-wolframic acid is amplified 25,000 times) of methicillin-resistant staphylococcus aureus (ATCC BAA-42) sterilization effect.
As shown in Figure 9, Control, control group, bacterium adds in the aforementioned nutrient solution, and 37 ℃, the 200rpm growth still presented normal morphology after 2 hours; Wild type colicin Ia, the same control group of bacterial growth condition added wild-type colicin Ia (50 μ g/ml) growth after two hours, and the bacterium form is still normal; AgrD, the same control group of bacterial growth condition added golden Portugal bacterium signal conduction polypeptide (10 μ g/ml) growth after two hours, and the bacterium form is still normal, and the thalline in upper end divides; Oxacillin, the same control group of bacterial growth condition added Oxazacillin (500 μ g/ml) growth after 2 hours, and the mycetocyte wall is the dissolving form; Ph-SA1, the same control group of bacterial growth condition, add anti-golden Portugal polypeptide (50 μ g/ml) growth half an hour after, bacterium is broken, content leaks, and remaining cell wall structures is complete.
Conclusion: use signal conduction polypeptide or wild-type colicin to handle merely and all can not kill MRSA, it is different fully with Oxazacillin that anti-golden Portugal polypeptide but can be killed MRSA and bactericidal mechanism.
The antibacterial activity in vitro of [embodiment 4] reorganization antimicrobial polypeptide
Below test is finished by institute of antibiotics, National Drug Administration Sichuan Pharmacology Lab.
One, experiment material
(1). medicine
1. ph-SA1: specification: 1.5mg/ml, lot number: PRE021101 is provided by West China medical college of Sichuan University.
2. cefazolin for injection: white powder, specification: the 0.5g/ bottle, lot number: 020418, Huabei Pharmaceutic Co., Ltd produces.
3. benzylpenicillin sodium for injection: white powder, specification: 800,000 unit/bottles, lot number: 00206604, Huabei Pharmaceutic Co., Ltd produces.
4. ampicillin for inj sodium: specification: the 1g/ bottle, lot number: F0203329, Huabei Pharmaceutic Co., Ltd produces.
5. oxacillin for inj: specification: the 0.5g/ bottle, lot number: F0203329, Huabei Pharmaceutic Co., Ltd produces.
6. vancomycin: specification: 2.5mg/ml, lot number: 020712, provide by the Drug Manufacturing Room of Sichuan Industrial Institute of Antibiotics.
Above sample sterilized water dissolved dilution, medicine final concentration are 128mg/L, 64mg/L, 32mg/L, 16mg/L, L8mg/, L4mg/L, 2mg/L, 1mg/L, 0.5mg/L, 0.25mg/L, 0.125mg/L, 0.06mg/L, 0.03mg/L, 0.015mg/L.
(2), bacterium
1. clinical isolates strain: the test bacterial strain uses therefor is 2002 from Sichuan, the Beijing area clinical separation pathogenic bacterium of collecting, all bacterial classifications are being collected isolating unit (clinical laboratory of Huaxi Medical Univ and Medical University Of Chongqing's contagious department Bacteriology Room) all after identifying, use after identify again with the API system this chamber again.
Golden yellow staphylococcus 168 strains (80 strains of MRSA bacterial strain, 88 strains of MSSA bacterial strain), staphylococcus epidermidis 46 strains (25 strains of MRSA bacterial strain, 21 strains of MSSE bacterial strain), faecalis 20 strains, Acinetobacter bauamnnii 20 strains, totally 274 strains.
2. standard Quality Control bacterial strain: streptococcus aureus ATCC25923, streptococcus aureus 29213 are that pharmacological room of Sichuan Industrial Institute of Antibiotics preserves bacterial strain.
(3), substratum
1. M-H substratum: Nat'l Pharmaceutical ﹠ Biological Products Control Institute's product, M-H broth culture: take by weighing 25g and add 100ml distilled water, heating for dissolving, packing, autoclaving, 116 ℃, 20 minutes.The M-H solid medium: take by weighing 36g, add 1000m distilled water, autoclaving, 20 minutes, is used for the drug sensitive test of gram-positive, negative aerophil by 116 ℃.
2. faecalis, suis are used blood meida, and it is formulated promptly to add 5-10% defiber rabbit blood in the M-H substratum, cultivate 24 hours for 37 ℃ among the 5%CO.
Two, test method
1. the mensuration of minimum inhibitory concentration MIC
Adopt the agar doubling dilution to measure the minimum inhibitory concentration (MIC) of ph-SA.With multiple spot inoculate instrument (Denley A400) with microbionation in the agar plate surface that contains different pharmaceutical concentration, every some bacteria containing amount is about 10 5CFU/ml is hatched 18～24 hours observationss for 37 ℃, is the minimum inhibitory concentration (MIC value) of medicine to this bacterium with the minimum concentration of contained drug in the no bacterial growth plate substratum.
Three, result
Experimental result sees Table 1.
The antibacterial activity in vitro of table 1 ph-SA1 relatively
Bacterial strain medicine MIC 50MIC 90The MIC scope
(strain number) (mg/L) (mg/L) (mg/L)
Streptococcus aureus ph-SA1 16＞16＜0.0005～64
MRSA bacterial strain Cephazolin＞128＞128 1～128
Penicillin 128＞128 1～128
Ampicillin Trihydrate 16 32
0.06～64
Oxazacillin＞128＞128 0.5～128
Vancomycin 0.25 1 0.004～2
Streptococcus aureus ph-SA1 0.5 16 0.125～＞64
MSSA bacterial strain Cephazolin 0.125 0.25
＜0.015～0.5
(88) penicillin 8 64
0.125～＞128
Ampicillin Trihydrate 0.25 1
＜0.015～0.25
Oxazacillin 0.125 0.125
＜0.015～0.25
Vancomycin 0.5 1
0.03～1
Staphylococcus epidermidis ph-SA1＞64＞64
0.25～＞64
MRSE bacterial strain Cephazolin 32＞128
0.06～＞128
(25) penicillin＞128＞128
8～＞128
Ampicillin Trihydrate 16 64
0.06～64
Oxazacillin 64＞128
0.5～＞128
Vancomycin 0.5 1
0.25～2
Staphylococcus epidermidis ph-SA1 0.25 16
0.125～32
MSSE bacterial strain Cephazolin 0.03 0.5
＜0.015～0.5
(21) penicillin 4 32
0.125～128
Ampicillin Trihydrate 0.03 0.25
＜0.015～0.25
Oxazacillin 0.03 0.125＜0.015～0.25
Vancomycin 0.5 0.5
0.06～0.5
Faecalis ph-SA1＞64＞64
8～＞64
(20) Cephazolin 32＞128
32～＞128
Penicillin 32 64
32～＞128
Ampicillin Trihydrate 0.5 16
0.25～16
Oxazacillin 16＞128
8～＞128
Vancomycin 12
0.5～50
Connect table 1
Bacterial strain medicine MIC 50MIC 90The MIC scope
(strain number) (mg/L) (mg/L) (mg/L)
Pseudomonas aeruginosa ph-SA1 16＞16 4～＞16
(20) Cephazolin＞128＞128 128～＞128
Penicillin＞128＞128＞128
Ampicillin Trihydrate＞128＞128 16～＞128
Oxazacillin＞128＞128＞128
Vancomycin＞128＞128＞128
Acinetobacter bauamnnii ph-SA1 32＞64 0.5～＞64
(20) Cephazolin＞128＞128＞128
Penicillin＞128＞128＞128
Ampicillin Trihydrate＞128＞128 16～＞128
Oxazacillin＞128＞128＞128
Vancomycin＞128＞128 128～＞128
The animal endogenous protective experiment of [embodiment 5] reorganization antimicrobial polypeptide
Below test is finished by institute of antibiotics, National Drug Administration Sichuan Pharmacology Lab.
One, experiment material
(1) medicine
The use medicine is the same.
Above sample sterilized water dissolved dilution, medicine injection final concentration is 10mg/kg, 5mg/gk, 2.5mg/kg.
(2) bacterium
1. standard Quality Control bacterial strain: streptococcus aureus ATCC BAA-42 (MRSA), faecalis ATCC700802 (VRE) are that pharmacological room of Sichuan Industrial Institute of Antibiotics preserves bacterial strain.
Two, experimental technique
170 of kunming mices, male and female half and half, body weight 18-22 gram, random packet is that the abdominal cavity is injected streptococcus aureus ATCC BAA-42 (MRSA) and two big groups of ATCC 700802 (VRE) respectively, wherein PH-SA1, penicillin, Ampicillin Trihydrate, Cephazolin and vancomycin test group are respectively 5 of every group of mouse, 10 of every group of mouse of control group.Behind the bacterium of abdominal injection lethal dose, each drug test group mouse respectively once from the medicine of intravenous injection 10mg, 5mg, 2.5mg/kg dosage, per 24 hours observationss, 7-14 days continuously, with the positive result of dead mouse.
Three, result
5mg, 3mg and 1mg drug dose group result show, Ph-SA1 group mouse survival rate has remained the constant gap with vancomycin group mouse survival rate in 3 dosage groups, Ph-SA1 is much better than vancomycin to the result of treatment that methicillin-resistant staphylococcus aureus resistance infects in therefore testing in vivo.This may be not only sterilization but also suppress due to the bacteriotoxin excretory double effects of Ph-SA1.Thereby this medicine may have the incomparable result of treatment of other antibiotic when clinical application.Other each antibiotic group mouse death rate 〉=80% has not possessed comparability; Control group is all dead in 48 hours.
Experimental result sees Table 2 and Figure 10.
Table 2 Ph-SA1 endogenous protective test-results (intravenously administrable)
Bacterium (bacterium number) infection dosage (cfu/ml) Medicine ??MIC ??(μg/ml) ????ED 50????mg/kg 95% fiducial limit (mg/kg) for the second time for the first time
Faecalis ATCC 200783 (8.0 * 10 6) ??Ph-SA1 ??4 ????4.23(3.70) ??2.90-7.62??????(2.78-5.09)
Kefzol ??＞128 ????＞10(12.91) ??＞10???????????(10.07-22.88)
Penicillin ??2 ????3.16(3.08) ??2.37-5.06??????(2.38-3.91)
The Ampicillin Trihydrate ??0.125 ????3.37(3.00) ??2.51-5.77??????(1.65-4.71)
Vancomycin ??16 ????3.17(2.64) ??2.54-4.14??????(2.10-3.39)
Staphylococcus aureus MRSA BAA-42 (5.5 * 10 5) ??Ph-SA1 ??0.5 ????4.32(4.23) ??3.28-6.18??????(3.05-5.86)
Kefzol ??0.5 ????9.04(8.78) ??7.04-17.54?????(6.84-12.16)
Penicillin ??8 ????＞10(14.71) ??＞10???????????(11.29-21.19)
The Ampicillin Trihydrate ??0.25 ????25.21(23.43) ??19.83-46.70????(16.87-36.58)
Vancomycin (0.08) ??0.06 ????3.64(3.53) ??2.75-6.16??????(2.66-4.73)
Staphylococcus aureus MRSA 02-18 (1.7 * 10 5) ??Ph-SA1 ??0.5 ????3.84(4.24) ??2.65-5.32??????(2.54-7.08)
Kefzol ??128 ????11.82(＞10) ??8.55-25.02?????(＞10)
Penicillin ??＞16 ????7.28(11.33) ??5.51-12.31?????(7.33-109.96)
The Ampicillin Trihydrate ??8 ????32.70(＞20) ??25.51-53.88????(＞20)
Vancomycin (0.077) ??0.06 ????2.27(2.39) ??1.75-2.97??????(1.89-3.07)
Staphylococcus aureus ATCC 29213 (3.6 * 10 5) ??Ph-SA1 ??0.25 ????10.35(8.87) ??8.04-15.38?????(7.00-11.85)
Kefzol ??0.03 ????29.03(11.20) ??17.82-61.64????(8.47-18.93)
Penicillin ??16 ????10.75(9.84) ??8.42-15.79?????(7.68-14.04)
The Ampicillin Trihydrate ??＜0.015 ????14.25(12.26) ??10.00-44.51????(9.80-18.29)
Vancomycin (0.2) ??0.5 ????2.32(2.09) ??1.71-3.29??????(1.57-2.76)
Colon bacillus 02-1-65 (3.3 * 10 5) ??Ph-SA1 ??1 ????3.14(3.12) ??2.48-4.28??????(2.49-4.12)
Kefzol ??＞128 ????＞30.77 ????(＞30.77) ??＞30.77????????(＞30.77)
Penicillin ??＞128 ????＞30.77 ????(＞30.77) ??＞30.77????????(＞30.77)
The Ampicillin Trihydrate ??＞128 ????＞30.77 ????(＞30.77) ??＞30.77????????(＞30.77)
Vancomycin ??128 ????＞30.77 ????(＞30.77) ????＞30.77??????(＞30.77)
Bacillus proteus 02321 (4.6 * 10 5) Ph-SA1 ??0.25 ????3.33(3.25) ????2.64-4.61????(2.64-4.17)
Kefzol ??2 ????＞30.77 ????＞30.77
Penicillin ??＞128 ????＞30.77 ????＞30.77
The Ampicillin Trihydrate ??2 ????＞30.77 ????＞30.77
Vancomycin ??＞128 ????＞56.02 ????＞30.77
By result in table 1 and the table 2 as can be seen:
1.MRSA antibacterial activity in vitro relatively: Oxazacillin MIC90 measured value is bigger 8 times than PH-SA1 measured value, and is bigger 128 times than vancomycin measured value.PH-SA1 molecular weight (70,0000) is 49 times of vancomycin molecular weight (1,400), 175 times of Oxazacillin molecular weight (400).Demarcate by same medicine molecule number in the unit volume, then PH-SA1 is equivalent to vancomycin 3 times to the MRSA antibacterial activity in vitro to the MRSA antibacterial activity in vitro, is equivalent to Oxazacillin 1,400 times to the MRSA antibacterial activity in vitro.
2.Ph-SA1 the responsive rate (is boundary with 16mg/L) to MSSA is 91%, responsive rate to MRSA is 77%, responsive rate to MSSE is 91%, responsive rate to MRSE is 40%, to enterococcal responsive rate is 30%, responsive rate to Pseudomonas aeruginosa is 50%, is 40% to the responsive rate of Acinetobacter bauamnnii.
3.SA in the infecting mouse endogenous protective preliminary experiment, the mortality ratio minimum (40%) of PH-SA1 group under minimizing agent dose (2.5mg/kg) condition, other is respectively organized mortality ratio and is respectively penicillin group 60%, ammonia click XiLin group 100%, vancomycin 60%, control group 100%.This result shows the protection ratio of PH-SA1 the highest (60%); The protection ratio of penicillin and vancomycin take second place (40%).
[embodiment 6] reorganization antimicrobial polypeptide is to the influence of epidermis aureus growth
Bacterium is a staphylococcus epidermidis reference culture of preserving center (State Pharmaceutical Administration's biological products assay institute, Beijing the Temple of Heaven) available from Chinese bacterial classification, numbering 26069, bacterium liquid 5 microlitres (10 8CFU/ml level bacterium amount) adds 1% Tryptones, 1%NaCl, 0.5% yeast, 0.5% glucose, 1%HK 2PO 4Nutrient solution 10ml in, prepare 4 groups altogether, first group adds 0.3M NaCl+50mM borate buffer (the antimicrobial polypeptide amount of liquid that adds in amount and the experimental group is identical) in contrast, second group adds 2 μ g/ml Benzylpenicillin sodiums, and surplus each experimental group respectively adds 2 μ g/ml antimicrobial polypeptide Ph-SA1 and Ph-SE (preserving liquid is 0.3M NaCl+50mM borate buffer).
The above-mentioned liquid of respectively organizing places the 100ml Erlenmeyer flask, 200rpm, 37 ℃ of growths, the 100 μ l that per hour sample add in the 96 hole enzyme plates through spectrophotometer (A 595nm) colorimetric bacteria tested growth turbidity, the growth curve of bacteria that draws comes the inhibitory effect of comparison antimicrobial polypeptide, the results are shown in Figure 11.
As seen from the figure, the growth of staphylococcus epidermidis can be suppressed by anti-golden Portugal polypeptide portion, can effectively be suppressed by anti-table Portugal polypeptide.
The comparison of [embodiment 7] several reorganization antimicrobial polypeptide fungistatic effects
Below the bacteriostatic activity of 6 kinds of antimicrobial polypeptides of the present invention's preparation has been compared in experiment, and bacterium is the USS bacterial strain, ATCC BAA-42 methicillin-resistant staphylococcus aureus, bacterium liquid 5 microlitres (10 8CFU/ml level bacterium amount) adds 1% Tryptones, 1%NaCl, 0.5% yeast, 0.5% glucose, 1%HK 2PO 4Nutrient solution 10ml in, prepare 8 groups altogether, first group adds 0.3M NaCl+50mM borate buffer (the antimicrobial polypeptide amount of liquid that adds in amount and the experimental group is identical) in contrast, second group adds 5 μ g/ml Benzylpenicillin sodiums, and surplus each experimental group respectively adds 5 μ g/ml antimicrobial polypeptide Ph-SA1, Ph-SA2, Ph-SA3, Ph-SA4, Ph-SE and Ph-SAR1 (preserving liquid is 0.3M NaCl+50mM borate buffer).
The above-mentioned liquid of respectively organizing places the 100ml Erlenmeyer flask, 200rpm, 37 ℃ of growths, the 100 μ l that per hour sample add in the 96 hole enzyme plates through spectrophotometer (A 595nm) colorimetric bacteria tested growth turbidity, the growth curve of bacteria that draws comes the inhibitory effect of comparison antimicrobial polypeptide, the results are shown in Figure 12.
The result shows that the growth of methicillin-resistant gold Portugal bacterium almost can not be suppressed by Oxazacillin, and the fungistatic effect of Ph-SAR1 is the poorest, and that the inhibition effect is best is Ph-SA1 and Ph-SA4, secondly is Ph-SE, and Ph-SA3 and Ph-SA2 take second place again.
Above detailed description of the present invention does not limit the present invention, and those skilled in the art can make various changes and distortion according to the present invention, only otherwise break away from spirit of the present invention, all should belong to the scope of claims of the present invention.
SEQUENCE?LISTING
＜110〉Yanghui Biotechnology Co Ltd, Chengdu
＜120〉a kind of novel antibacterial polypeptide and preparation method thereof
<130>1552
<160>33
<170>PatentIn?version?3.1
<210>1
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<221>CDS
<222>(1)..(24)
＜223〉coding streptococcus aureus Agr D I signal conduction polypeptide
<400>1
tat?tcc?acc?tgt?gat?ttt?ata?atg????????????????????????????????????????24
Tyr?Ser?Thr?Cys?Asp?Phe?Ile?Met
1???????????????5
<210>2
<211>8
<212>PRT
＜213〉artificial sequence
<400>2
Tyr?Ser?Thr?Cys?Asp?Phe?Ile?Met
1???????????????5
<210>3
<211>27
<212>DNA
＜213〉artificial sequence
<220>
<221>CDS
<222>(1)..(27)
＜223〉coding streptococcus aureus Agr D II signal conduction polypeptide
<400>3
gga?gtt?aac?gca?tgt?tct?tcc?ctg?ttt???????????????????????????????????27
Gly?Val?Asn?Ala?Cys?Ser?Ser?Leu?Phe
1???????????????5
<210>4
<211>9
<212>PRT
＜213〉artificial sequence
<400>4
Gly?Val?Asn?Ala?Cys?Ser?Ser?Leu?Phe
1???????????????5
<210>5
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<221>CDS
<222>(1)..(24)
＜223〉coding streptococcus aureus Agr D III signal conduction polypeptide
<400>5
tat?ata?aac?tgt?gat?ttt?ctt?ctg??????????????????????????????????????24
Tyr?Ile?Asn?Cys?Asp?Phe?Leu?Leu
1???????????????5
<210>6
<211>8
<212>PRT
＜213〉artificial sequence
<400>6
Tyr?Ile?Asn?Cys?Asp?Phe?Leu?Leu
1???????????????5
<210>7
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<221>CDS
<222>(1)..(24)
＜223〉coding streptococcus aureus Agr D IV signal conduction polypeptide
<400>7
tat?tcc?acc?tgt?tac?ttt?ata?atg??????????????????????????????????????24
Tyr?Ser?Thr?Cys?Tyr?Phe?Ile?Met
1???????????????5
<210>8
<211>8
<212>PRT
＜213〉artificial sequence
<400>8
Tyr?Ser?Thr?Cys?Tyr?Phe?Ile?Met
1???????????????5
<210>9
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<221>CDS
<222>(1)..(24)
＜223〉coding staphylococcus epidermidis signal conduction polypeptide
<400>9
gat?tcc?gtt?tgt?gca?tcc?tat?ttt????????????????????????????????????????24
Asp?Ser?Val?Cys?Ala?Ser?Tyr?Phe
1???????????????5
<210>10
<211>8
<212>PRT
＜213〉artificial sequence
<400>10
Asp?Ser?Val?Cys?Ala?Ser?Tyr?Phe
1???????????????5
<210>11
<211>1878
<212>DNA
<213>Escherichia?coli
<220>
<221>misc_feature
<222>(1)..(1878)
＜223〉colicin gene
<400>11
atgtctgacc?ctgtacgtat?tacaaatccc?ggtgcagaat?cgctggggta?tgattcagat????60
ggccatgaaa?ttatggccgt?tgatatttat?gtaaaccctc?cacgtgtcga?tgtctttcat???120
ggtaccccgc?ctgcatggag?ttccttcggg?aacaaaacca?tctggggcgg?aaacgagtgg???180
gttgatgatt?ccccaacccg?aagtgatatc?gaaaaaaggg?acaaggaaat?cacagcgtac???240
aaaaacacgc?tcagcgcgca?gcagaaagag?aatgagaata?agcgtactga?agccggaaaa???300
cgcctctctg?cggcgattgc?tgcaagggaa?aaagatgaaa?acacactgaa?aacactccgt???360
gccggaaacg?cagatgccgc?tgatattaca?cgacaggagt?tcagactcct?gcaggcagag???420
ctgagagaat?acggattccg?tactgaaatc?gccggatatg?acgccctccg?gctgcataca???480
gagagccgga?tgctgtttgc?tgatgctgat?tctcttcgta?tatctccccg?ggaggccagg???540
tcgttaatcg?aacaggctga?aaaacggcag?aaggatgcgc?agaacgcaga?caagaaggcc???600
gctgatatgc?ttgctgaata?cgagcgcaga?aaaggtattc?tggacacccg?gttgtcagag???660
ctggaaaaaa?atggcggggc?agcccttgcc?gttcttgatg?cacaacaggc?ccgtctgctc???720
gggcagcaga?cacggaatga?cagggccatt?tcagaggccc?ggaataaact?cagttcagtg???780
acggaatcgc?ttaacacggc?ccgtaatgca?ttaaccagag?ctgaacaaca?gctgacgcaa???840
cagaaaaaca?cgcctgacgg?caaaacgata?gtttcccctg?aaaaattccc?ggggcgttca???900
tcaacaaatc?attctattgt?tgtgagcggt?gatccgagat?ttgccggtac?gataaaaatc???960
acaaccagcg?cagtcatcga?taaccgtgca?aacctgaatt?atcttctgag?ccattccggt??1020
ctggactata?aacgcaatat?tctgaatgac?cggaatccgg?tggtgacaga?ggatgtggaa??1080
ggtgacaaga?aaatttataa?tgctgaagtt?gctgaatggg?ataagttacg?gcaaagattg??1140
cttgatgcca?gaaataaaat?cacctctgct?gaatctgcgg?taaattcggc?gagaaataac??1200
ctcagtgcca?gaacaaatga?gcaaaagcat?gcaaatgacg?ctcttaatgc?cctgttgaag??1260
gaaaaagaga?atatacgtaa?ccagctttcc?ggcatcaatc?agaagatagc?ggaagagaaa??1320
agaaaacagg?atgaactgaa?ggcaacgaaa?gacgcaatta?atttcacaac?agagttcctg??1380
aaatcagttt?cagaaaaata?tggtgcaaaa?gctgagcagt?tagccagaga?gatggccggg??1440
caggctaaag?ggaagaaaat?acgtaatgtt?gaagaggcat?taaaaacgta?tgaaaagtac??1500
cgggctgaca?ttaacaaaaa?aattaatgca?aaagatcgtg?cagcgattgc?cgcagccctt??1560
gagtctgtga?agctgtctga?tatatcgtct?aatctgaaca?gattcagtcg?gggactggga??1620
tatgcaggaa?aatttacaag?tcttgctgac?tggatcactg?agtttggtaa?ggctgtccgg??1680
acagagaact?ggcgtcctct?ttttgttaaa?acagaaacca?tcatagcagg?caatgccgca??1740
acggctcttg?tggcactggt?cttcagtatt?cttaccggaa?gcgctttagg?cattatcggg??1800
tatggtttac?tgatggctgt?caccggtgcg?ctgattgatg?aatcgcttgt?ggaaaaagcg??1860
aataagttct?ggggtatt????????????????????????????????????????????????1878
<210>12
<211>1905
<212>DNA
＜213〉artificial sequence
<220>
<221>CDS
<222>(4)..(1902)
＜223〉coding staphylococcus polypeptide
<400>12
atg?tct?gac?cct?gta?cgt?att?aca?aat?ccc?ggt?gca?gaa?tcg?ctg?ggg???????48
Ser?Asp?Pro?Val?Arg?Ile?Thr?Asn?Pro?Gly?Ala?Glu?Ser?Leu?Gly
1???????????????5???????????????????10??????????????????15
tat?gat?tca?gat?ggc?cat?gaa?att?atg?gcc?gtt?gat?att?tat?gta?aac???????96
Tyr?Asp?Ser?Asp?Gly?His?Glu?Ile?Met?Ala?Val?Asp?Ile?Tyr?Val?Asn
20??????????????????25??????????????????30
cct?cca?cgt?gtc?gat?gtc?ttt?cat?ggt?acc?ccg?cct?gca?tgg?agt?tcc??????144
Pro?Pro?Arg?Val?Asp?Val?Phe?His?Gly?Thr?Pro?Pro?Ala?Trp?Ser?Ser
35??????????????????40??????????????????45
ttc?ggg?aac?aaa?acc?atc?tgg?ggc?gga?aac?gag?tgg?gtt?gat?gat?tcc??????192
Phe?Gly?Asn?Lys?Thr?Ile?Trp?Gly?Gly?Asn?Glu?Trp?Val?Asp?Asp?Ser
50??????????????????55??????????????????60
cca?acc?cga?agt?gat?atc?gaa?aaa?agg?gac?aag?gaa?atc?aca?gcg?tac??????240
Pro?Thr?Arg?Ser?Asp?Ile?Glu?Lys?Arg?Asp?Lys?Glu?Ile?Thr?Ala?Tyr
65??????????????????70??????????????????75
aaa?aac?acg?ctc?agc?gcg?cag?cag?aaa?gag?aat?gag?aat?aag?cgt?act??????288
Lys?Asn?Thr?Leu?Ser?Ala?Gln?Gln?Lys?Glu?Asn?Glu?Asn?Lys?Arg?Thr
80??????????????????85??????????????????90??????????????????95
gaa?gcc?gga?aaa?cgc?ctc?tct?gcg?gcg?att?gct?gca?agg?gaa?aaa?gat??????336
Glu?Ala?Gly?Lys?Arg?Leu?Ser?Ala?Ala?Ile?Ala?Ala?Arg?Glu?Lys?Asp
100?????????????????105?????????????????110
gaa?aac?aca?ctg?aaa?aca?ctc?cgt?gcc?gga?aac?gca?gat?gcc?gct?gat??????384
Glu?Asn?Thr?Leu?Lys?Thr?Leu?Arg?Ala?Gly?Asn?Ala?Asp?Ala?Ala?Asp
115?????????????????120?????????????????125
att?aca?cga?cag?gag?ttc?aga?ctc?ctg?cag?gca?gag?ctg?aga?gaa?tac??????432
Ile?Thr?Arg?Gln?Glu?Phe?Arg?Leu?Leu?Gln?Ala?Glu?Leu?Arg?Glu?Tyr
130?????????????????135?????????????????140
gga?ttc?cgt?act?gaa?atc?gcc?gga?tat?gac?gcc?ctc?cgg?ctg?cat?aca??????480
Gly?Phe?Arg?Thr?Glu?Ile?Ala?Gly?Tyr?Asp?Ala?Leu?Arg?Leu?His?Thr
145?????????????????150?????????????????155
gag?agc?cgg?atg?ctg?ttt?gct?gat?gct?gat?tct?ctt?cgt?ata?tct?ccc??????528
Glu?Ser?Arg?Met?Leu?Phe?Ala?Asp?Ala?Asp?Ser?Leu?Arg?Ile?Ser?Pro
160?????????????????165?????????????????170?????????????????175
cgg?gag?gcc?agg?tcg?tta?atc?gaa?cag?gct?gaa?aaa?cgg?cag?aag?gat??????576
Arg?Glu?Ala?Arg?Ser?Leu?Ile?Glu?Gln?Ala?Glu?Lys?Arg?Gln?Lys?Asp
180?????????????????185?????????????????190
gcg?cag?aac?gca?gac?aag?aag?gcc?gct?gat?atg?ctt?gct?gaa?tac?gag??????624
Ala?Gln?Asn?Ala?Asp?Lys?Lys?Ala?Ala?Asp?Met?Leu?Ala?Glu?Tyr?Glu
195?????????????????200?????????????????205
cgc?aga?aaa?ggt?att?ctg?gac?acc?cgg?ttg?tca?gag?ctg?gaa?aaa?aat??????672
Arg?Arg?Lys?Gly?Ile?Leu?Asp?Thr?Arg?Leu?Ser?Glu?Leu?Glu?Lys?Asn
210?????????????????215?????????????????220
ggc?ggg?gca?gcc?ctt?gcc?gtt?ctt?gat?gca?caa?cag?gcc?cgt?ctg?ctc??????720
Gly?Gly?Ala?Ala?Leu?Ala?Val?Leu?Asp?Ala?Gln?Gln?Ala?Arg?Leu?Leu
225?????????????????230?????????????????235
ggg?cag?cag?aca?cgg?aat?gac?agg?gcc?att?tca?gag?gcc?cgg?aat?aaa??????768
Gly?Gln?Gln?Thr?Arg?Asn?Asp?Arg?Ala?Ile?Ser?Glu?Ala?Arg?Asn?Lys
240?????????????????245?????????????????250?????????????????255
ctc?agt?tca?gtg?acg?gaa?tcg?ctt?aac?acg?gcc?cgt?aat?gca?tta?acc??????816
Leu?Ser?Ser?Val?Thr?Glu?Ser?Leu?Asn?Thr?Ala?Arg?Asn?Ala?Leu?Thr
260?????????????????265?????????????????270
aga?gct?gaa?caa?cag?ctg?acg?caa?cag?aaa?aac?acg?cct?gac?ggc?aaa??????864
Arg?Ala?Glu?Gln?Gln?Leu?Thr?Gln?Gln?Lys?Asn?Thr?Pro?Asp?Gly?Lys
275?????????????????280?????????????????285
acg?ata?gtt?tcc?cct?gaa?aaa?ttc?ccg?ggg?cgt?tca?tca?aca?aat?cat??????912
Thr?Ile?Val?Ser?Pro?Glu?Lys?Phe?Pro?Gly?Arg?Ser?Ser?Thr?Asn?His
290?????????????????295?????????????????300
tct?att?gtt?gtg?agc?ggt?gat?ccg?aga?ttt?gcc?ggt?acg?ata?aaa?atc??????960
Ser?Ile?Val?Val?Ser?Gly?Asp?Pro?Arg?Phe?Ala?Gly?Thr?Ile?Lys?Ile
305?????????????????310?????????????????315
aca?acc?agc?gca?gtc?atc?gat?aac?cgt?gca?aac?ctg?aat?tat?ctt?ctg?????1008
Thr?Thr?Ser?Ala?Val?Ile?Asp?Asn?Arg?Ala?Asn?Leu?Asn?Tyr?Leu?Leu
320?????????????????325?????????????????330????????????????335
agc?cat?tcc?ggt?ctg?gac?tat?aaa?cgc?aat?att?ctg?aat?gac?cgg?aat?????1056
Ser?His?Ser?Gly?Leu?Asp?Tyr?Lys?Arg?Asn?Ile?Leu?Asn?Asp?Arg?Asn
340?????????????????345?????????????????350
ccg?gtg?gtg?aca?gag?gat?gtg?gaa?ggt?gac?aag?aaa?att?tat?aat?gct?????1104
Pro?Val?Val?Thr?Glu?Asp?Val?Glu?Gly?Asp?Lys?Lys?Ile?Tyr?Asn?Ala
355?????????????????360?????????????????365
gaa?gtt?gct?gaa?tgg?gat?aag?tta?cgg?caa?aga?ttg?ctt?gat?gcc?aga?????1152
Glu?Val?Ala?Glu?Trp?Asp?Lys?Leu?Arg?Gln?Arg?Leu?Leu?Asp?Ala?Arg
370?????????????????375?????????????????380
aat?aaa?atc?acc?tct?gct?gaa?tct?gcg?gta?aat?tcg?gcg?aga?aat?aac?????1200
Asn?Lys?Ile?Thr?Ser?Ala?Glu?Ser?Ala?Val?Asn?Ser?Ala?Arg?Asn?Asn
385?????????????????390?????????????????395
ctc?agt?gcc?aga?aca?aat?gag?caa?aag?cat?gca?aat?gac?gct?ctt?aat?????1248
Leu?Ser?Ala?Arg?Thr?Asn?Glu?Gln?Lys?His?Ala?Asn?Asp?Ala?Leu?Asn
400?????????????????405?????????????????410?????????????????415
gcc?ctg?ttg?aag?gaa?aaa?gag?aat?ata?cgt?aac?cag?ctt?tcc?ggc?atc?????1296
Ala?Leu?Leu?Lys?Glu?Lys?Glu?Asn?Ile?Arg?Asn?Gln?Leu?Ser?Gly?Ile
420?????????????????425?????????????????430
aat?cag?aag?ata?gcg?gaa?gag?aaa?aga?aaa?cag?gat?gaa?ctg?aag?gca?????1344
Asn?Gln?Lys?Ile?Ala?Glu?Glu?Lys?Arg?Lys?Gln?Asp?Glu?Leu?Lys?Ala
435?????????????????440?????????????????445
acg?aaa?gac?gca?att?aat?ttc?aca?aca?gag?ttc?ctg?aaa?tca?gtt?tca?????1392
Thr?Lys?Asp?Ala?Ile?Asn?Phe?Thr?Thr?Glu?Phe?Leu?Lys?Ser?Val?Ser
450?????????????????455?????????????????460
gaa?aaa?tat?ggt?gca?aaa?gct?gag?cag?tta?gcc?aga?gag?atg?gcc?ggg?????1440
Glu?Lys?Tyr?Gly?Ala?Lys?Ala?Glu?Gln?Leu?Ala?Arg?Glu?Met?Ala?Gly
465?????????????????470?????????????????475
cag?gct?aaa?ggg?aag?aaa?ata?cgt?aat?gtt?gaa?gag?gca?tta?aaa?acg?????1488
Gln?Ala?Lys?Gly?Lys?Lys?Ile?Arg?Asn?Val?Glu?Glu?Ala?Leu?Lys?Thr
480?????????????????485?????????????????490?????????????????495
tat?gaa?aag?tac?cgg?gct?gac?att?aac?aaa?aaa?att?aat?gca?aaa?gat?????1536
Tyr?Glu?Lys?Tyr?Arg?Ala?Asp?Ile?Asn?Lys?Lys?Ile?Asn?Ala?Lys?Asp
500?????????????????505?????????????????510
cgt?gca?gcg?att?gcc?gca?gcc?ctt?gag?tct?gtg?aag?ctg?tct?gat?ata?????1584
Arg?Ala?Ala?Ile?Ala?Ala?Ala?Leu?Glu?Ser?Val?Lys?Leu?Ser?Asp?Ile
515?????????????????520?????????????????525
tcg?tct?aat?ctg?aac?aga?ttc?agt?cgg?gga?ctg?gga?tat?gca?gga?aaa?????1632
Ser?Ser?Asn?Leu?Asn?Arg?Phe?Ser?Arg?Gly?Leu?Gly?Tyr?Ala?Gly?Lys
530?????????????????535?????????????????540
ttt?aca?agt?ctt?gct?gac?tgg?atc?act?gag?ttt?ggt?aag?gct?gtc?cgg?????1680
Phe?Thr?Ser?Leu?Ala?Asp?Trp?Ile?Thr?Glu?Phe?Gly?Lys?Ala?Val?Arg
545?????????????????550?????????????????555
aca?gag?aac?tgg?cgt?cct?ctt?ttt?gtt?aaa?aca?gaa?acc?atc?ata?gca?????1728
Thr?Glu?Asn?Trp?Arg?Pro?Leu?Phe?Val?Lys?Thr?Glu?Thr?Ile?Ile?Ala
560?????????????????565?????????????????570?????????????????575
ggc?aat?gcc?gca?acg?gct?ctt?gtg?gca?ctg?gtc?ttc?agt?att?ctt?acc?????1776
Gly?Asn?Ala?Ala?Thr?Ala?Leu?Val?Ala?Leu?Val?Phe?Ser?Ile?Leu?Thr
580?????????????????585?????????????????590
gga?agc?gct?tta?ggc?att?atc?ggg?tat?ggt?tta?ctg?atg?gct?gtc?acc?????1824
Gly?Ser?Ala?Leu?Gly?Ile?Ile?Gly?Tyr?Gly?Leu?Leu?Met?Ala?Val?Thr
595?????????????????600?????????????????605
ggt?gcg?ctg?att?gat?gaa?tcg?ctt?gtg?gaa?aaa?gcg?aat?aag?ttc?tgg?????1872
Gly?Ala?Leu?Ile?Asp?Glu?Ser?Leu?Val?Glu?Lys?Ala?Asn?Lys?Phe?Trp
610?????????????????615?????????????????620
ggt?att?tat?tcc?acc?tgt?gat?ttt?ata?atg?taa?????????????????????????1905
Gly?Ile?Tyr?Ser?Thr?Cys?Asp?Phe?Ile?Met
625?????????????????630
<210>13
<211>633
<212>PRT
＜213〉artificial sequence
<400>13
Ser?Asp?Pro?Val?Arg?Ile?Thr?Asn?Pro?Gly?Ala?Glu?Ser?Leu?Gly?Tyr
1???????????????5???????????????????10??????????????????15
Asp?Ser?Asp?Gly?His?Glu?Ile?Met?Ala?Val?Asp?Ile?Tyr?Val?Asn?Pro
20??????????????????25??????????????????30
Pro?Arg?Val?Asp?Val?Phe?His?Gly?Thr?Pro?Pro?Ala?Trp?Ser?Ser?Phe
35??????????????????40??????????????????45
Gly?Asn?Lys?Thr?Ile?Trp?Gly?Gly?Asn?Glu?Trp?Val?Asp?Asp?Ser?Pro
50??????????????????55??????????????????60
Thr?Arg?Ser?Asp?Ile?Glu?Lys?Arg?Asp?Lys?Glu?Ile?Thr?Ala?Tyr?Lys
65??????????????????70??????????????????75??????????????????80
Asn?Thr?Leu?Ser?Ala?Gln?Gln?Lys?Glu?Asn?Glu?Asn?Lys?Arg?Thr?Glu
85??????????????????90??????????????????95
Ala?Gly?Lys?Arg?Leu?Ser?Ala?Ala?Ile?Ala?Ala?Arg?Glu?Lys?Asp?Glu
100?????????????????105?????????????????110
Asn?Thr?Leu?Lys?Thr?Leu?Arg?Ala?Gly?Asn?Ala?Asp?Ala?Ala?Asp?Ile
115?????????????????120??????????????????125
Thr?Arg?Gln?Glu?Phe?Arg?Leu?Leu?Gln?Ala?Glu?Leu?Arg?Glu?Tyr?Gly
130?????????????????135?????????????????140
Phe?Arg?Thr?Glu?Ile?Ala?Gly?Tyr?Asp?Ala?Leu?Arg?Leu?His?Thr?Glu
145?????????????????150?????????????????155?????????????????160
Ser?Arg?Met?Leu?Phe?Ala?Asp?Ala?Asp?Ser?Leu?Arg?Ile?Ser?Pro?Arg
165?????????????????170?????????????????175
Glu?Ala?Arg?Ser?Leu?Ile?Glu?Gln?Ala?Glu?Lys?Arg?Gln?Lys?Asp?Ala
180?????????????????185?????????????????190
Gln?Asn?Ala?Asp?Lys?Lys?Ala?Ala?Asp?Met?Leu?Ala?Glu?Tyr?Glu?Arg
195?????????????????200?????????????????205
Arg?Lys?Gly?Ile?Leu?Asp?Thr?Arg?Leu?Ser?Glu?Leu?Glu?Lys?Asn?Gly
210?????????????????215?????????????????220
Gly?Ala?Ala?Leu?Ala?Val?Leu?Asp?Ala?Gln?Gln?Ala?Arg?Leu?Leu?Gly
225?????????????????230?????????????????235?????????????????240
Gln?Gln?Thr?Arg?Asn?Asp?Arg?Ala?Ile?Ser?Glu?Ala?Arg?Asn?Lys?Leu
245?????????????????250?????????????????255
Ser?Ser?Val?Thr?Glu?Ser?Leu?Asn?Thr?Ala?Arg?Asn?Ala?Leu?Thr?Arg
260?????????????????265??????????????????270
Ala?Glu?Gln?Gln?Leu?Thr?Gln?Gln?Lys?Asn?Thr?Pro?Asp?Gly?Lys?Thr
275????????????????280?????????????????285
Ile?Val?Ser?Pro?Glu?Lys?Phe?Pro?Gly?Arg?Ser?Ser?Thr?Asn?His?Ser
290?????????????????295?????????????????300
Ile?Val?Val?Ser?Gly?Asp?Pro?Arg?Phe?Ala?Gly?Thr?Ile?Lys?Ile?Thr
305?????????????????310?????????????????315?????????????????320
Thr?Ser?Ala?Val?Ile?Asp?Asn?Arg?Ala?Asn?Leu?Asn?Tyr?Leu?Leu?Ser
325?????????????????330?????????????????335
His?Ser?Gly?Leu?Asp?Tyr?Lys?Arg?Asn?Ile?Leu?Asn?Asp?Arg?Asn?Pro
340?????????????????345?????????????????350
Val?Val?Thr?Glu?Asp?Val?Glu?Gly?Asp?Lys?Lys?Ile?Tyr?Asn?Ala?Glu
355?????????????????360?????????????????365
Val?Ala?Glu?Trp?Asp?Lys?Leu?Arg?Gln?Arg?Leu?Leu?Asp?Ala?Arg?Asn
370?????????????????375?????????????????380
Lys?Ile?Thr?Ser?Ala?Glu?Ser?Ala?Val?Asn?Ser?Ala?Arg?Asn?Asn?Leu
385?????????????????390?????????????????395?????????????????400
Ser?Ala?Arg?Thr?Asn?Glu?Gln?Lys?His?Ala?Asn?Asp?Ala?Leu?Asn?Ala
405?????????????????410?????????????????415
Leu?Leu?Lys?Glu?Lys?Glu?Asn?Ile?Arg?Asn?Gln?Leu?Ser?Gly?Ile?Asn
420?????????????????425?????????????????430
Gln?Lys?Ile?Ala?Glu?Glu?Lys?Arg?Lys?Gln?Asp?Glu?Leu?Lys?Ala?Thr
435?????????????????440?????????????????445
Lys?Asp?Ala?Ile?Asn?Phe?Thr?Thr?Glu?Phe?Leu?Lys?Ser?Val?Ser?Glu
450?????????????????455?????????????????460
Lys?Tyr?Gly?Ala?Lys?Ala?Glu?Gln?Leu?Ala?Arg?Glu?Met?Ala?Gly?Gln
465?????????????????470?????????????????475?????????????????480
Ala?Lys?Gly?Lys?Lys?Ile?Arg?Asn?Val?Glu?Glu?Ala?Leu?Lys?Thr?Tyr
485?????????????????490?????????????????495
Glu?Lys?Tyr?Arg?Ala?Asp?Ile?Asn?Lys?Lys?Ile?Asn?Ala?Lys?Asp?Arg
500????????????????505?????????????????510
Ala?Ala?Ile?Ala?Ala?Ala?Leu?Glu?Ser?Val?Lys?Leu?Ser?Asp?Ile?Ser
515?????????????????520?????????????????525
Ser?Asn?Leu?Asn?Arg?Phe?Ser?Arg?Gly?Leu?Gly?Tyr?Ala?Gly?Lys?Phe
530?????????????????535?????????????????540
Thr?Ser?Leu?Ala?Asp?Trp?Ile?Thr?Glu?Phe?Gly?Lys?Ala?Val?Arg?Thr
545?????????????????550?????????????????555?????????????????560
Glu?Asn?Trp?Arg?Pro?Leu?Phe?Val?Lys?Thr?Glu?Thr?Ile?Ile?Ala?Gly
565??????????????????570????????????????575
Asn?Ala?Ala?Thr?Ala?Leu?Val?Ala?Leu?Val?Phe?Ser?Ile?Leu?Thr?Gly
580?????????????????585?????????????????590
Ser?Ala?Leu?Gly?Ile?Ile?Gly?Tyr?Gly?Leu?Leu?Met?Ala?Val?Thr?Gly
595?????????????????600?????????????????605
Ala?Leu?Ile?Asp?Glu?Ser?Leu?Val?Glu?Lys?Ala?Asn?Lys?Phe?Trp?Gly
610?????????????????615?????????????????620
Ile?Tyr?Ser?Thr?Cys?Asp?Phe?Ile?Met
625?????????????????630
<210>14
<211>1908
<212>DNA
＜213〉artificial sequence
<220>
<221>CDS
<222>(4)..(1905)
＜223〉coding staphylococcus polypeptide
<400>14
atg?tct?gac?cct?gta?cgt?att?aca?aat?ccc?ggt?gca?gaa?tcg?ctg?ggg?????48
Ser?Asp?Pro?Val?Arg?Ile?Thr?Asn?Pro?Gly?Ala?Glu?Ser?Leu?Gly
1???????????????5???????????????????10??????????????????15
tat?gat?tca?gat?ggc?cat?gaa?att?atg?gcc?gtt?gat?att?tat?gta?aac?????96
Tyr?Asp?Ser?Asp?Gly?His?Glu?Ile?Met?Ala?Val?Asp?Ile?Tyr?Val?Asn
20??????????????????25??????????????????30
cct?cca?cgt?gtc?gat?gtc?ttt?cat?ggt?acc?ccg?cct?gca?tgg?agt?tcc????144
Pro?Pro?Arg?Val?Asp?Val?Phe?His?Gly?Thr?Pro?Pro?Ala?Trp?Ser?Ser
35??????????????????40??????????????????45
ttc?ggg?aac?aaa?acc?atc?tgg?ggc?gga?aac?gag?tgg?gtt?gat?gat?tcc????192
Phe?Gly?Asn?Lys?Thr?Ile?Trp?Gly?Gly?Asn?Glu?Trp?Val?Asp?Asp?Ser
50??????????????????55??????????????????60
cca?acc?cga?agt?gat?atc?gaa?aaa?agg?gac?aag?gaa?atc?aca?gcg?tac????240
Pro?Thr?Arg?Ser?Asp?Ile?Glu?Lys?Arg?Asp?Lys?Glu?Ile?Thr?Ala?Tyr
65??????????????????70??????????????????75
aaa?aac?acg?ctc?agc?gcg?cag?cag?aaa?gag?aat?gag?aat?aag?cgt?act????288
Lys?Asn?Thr?Leu?Ser?Ala?Gln?Gln?Lys?Glu?Asn?Glu?Asn?Lys?Arg?Thr
80??????????????????85??????????????????90??????????????????95
gaa?gcc?gga?aaa?cgc?ctc?tct?gcg?gcg?att?gct?gca?agg?gaa?aaa?gat????336
Glu?Ala?Gly?Lys?Arg?Leu?Ser?Ala?Ala?Ile?Ala?Ala?Arg?Glu?Lys?Asp
100?????????????????105?????????????????110
gaa?aac?aca?ctg?aaa?aca?ctc?cgt?gcc?gga?aac?gca?gat?gcc?gct?gat????384
Glu?Asn?Thr?Leu?Lys?Thr?Leu?Arg?Ala?Gly?Asn?Ala?Asp?Ala?Ala?Asp
115?????????????????120?????????????????125
att?aca?cga?cag?gag?ttc?aga?ctc?ctg?cag?gca?gag?ctg?aga?gaa?tac????432
Ile?Thr?Arg?Gln?Glu?Phe?Arg?Leu?Leu?Gln?Ala?Glu?Leu?Arg?Glu?Tyr
130?????????????????135?????????????????140
gga?ttc?cgt?act?gaa?atc?gcc?gga?tat?gac?gcc?ctc?cgg?ctg?cat?aca?????480
Gly?Phe?Arg?Thr?Glu?Ile?Ala?Gly?Tyr?Asp?Ala?Leu?Arg?Leu?His?Thr
145?????????????????150?????????????????155
gag?agc?cgg?atg?ctg?ttt?gct?gat?gct?gat?tct?ctt?cgt?ata?tct?ccc?????528
Glu?Ser?Arg?Met?Leu?Phe?Ala?Asp?Ala?Asp?Ser?Leu?Arg?Ile?Ser?Pro
160?????????????????165?????????????????170?????????????????175
cgg?gag?gcc?agg?tcg?tta?atc?gaa?cag?gct?gaa?aaa?cgg?cag?aag?gat?????576
Arg?Glu?Ala?Arg?Ser?Leu?Ile?Glu?Gln?Ala?Glu?Lys?Arg?Gln?Lys?Asp
180?????????????????185?????????????????190
gcg?cag?aac?gca?gac?aag?aag?gcc?gct?gat?atg?ctt?gct?gaa?tac?gag?????624
Ala?Gln?Asn?Ala?Asp?Lys?Lys?Ala?Ala?Asp?Met?Leu?Ala?Glu?Tyr?Glu
195?????????????????200?????????????????205
cgc?aga?aaa?ggt?att?ctg?gac?acc?cgg?ttg?tca?gag?ctg?gaa?aaa?aat?????672
Arg?Arg?Lys?Gly?Ile?Leu?Asp?Thr?Arg?Leu?Ser?Glu?Leu?Glu?Lys?Asn
210?????????????????215?????????????????220
ggc?ggg?gca?gcc?ctt?gcc?gtt?ctt?gat?gca?caa?cag?gcc?cgt?ctg?ctc?????720
Gly?Gly?Ala?Ala?Leu?Ala?Val?Leu?Asp?Ala?Gln?Gln?Ala?Arg?Leu?Leu
225?????????????????230?????????????????235
ggg?cag?cag?aca?cgg?aat?gac?agg?gcc?att?tca?gag?gcc?cgg?aat?aaa?????768
Gly?Gln?Gln?Thr?Arg?Asn?Asp?Arg?Ala?Ile?Ser?Glu?Ala?Arg?Asn?Lys
240?????????????????245?????????????????250?????????????????255
ctc?agt?tca?gtg?acg?gaa?tcg?ctt?aac?acg?gcc?cgt?aat?gca?tta?acc?????816
Leu?Ser?Ser?Val?Thr?Glu?Ser?Leu?Asn?Thr?Ala?Arg?Asn?Ala?Leu?Thr
260?????????????????265?????????????????270
aga?gct?gaa?caa?cag?ctg?acg?caa?cag?aaa?aac?acg?cct?gac?ggc?aaa?????864
Arg?Ala?Glu?Gln?Gln?Leu?Thr?Gln?Gln?Lys?Asn?Thr?Pro?Asp?Gly?Lys
275?????????????????280?????????????????285
acg?ata?gtt?tcc?cct?gaa?aaa?ttc?ccg?ggg?cgt?tca?tca?aca?aat?cat?????912
Thr?Ile?Val?Ser?Pro?Glu?Lys?Phe?Pro?Gly?Arg?Ser?Ser?Thr?Asn?His
290?????????????????295?????????????????300
tct?att?gtt?gtg?agc?ggt?gat?ccg?aga?ttt?gcc?ggt?acg?ata?aaa?atc?????960
Ser?Ile?Val?Val?Ser?Gly?Asp?Pro?Arg?Phe?Ala?Gly?Thr?Ile?Lys?Ile
305?????????????????310?????????????????315
aca?acc?agc?gca?gtc?atc?gat?aac?cgt?gca?aac?ctg?aat?tat?ctt?ctg????1008
Thr?Thr?Ser?Ala?Val?Ile?Asp?Asn?Arg?Ala?Asn?Leu?Asn?Tyr?Leu?Leu
320?????????????????325?????????????????330?????????????????335
agc?cat?tcc?ggt?ctg?gac?tat?aaa?cgc?aat?att?ctg?aat?gac?cgg?aat????1056
Ser?His?Ser?Gly?Leu?Asp?Tyr?Lys?Arg?Asn?Ile?Leu?Asn?Asp?Arg?Asn
340?????????????????345?????????????????350
ccg?gtg?gtg?aca?gag?gat?gtg?gaa?ggt?gac?aag?aaa?att?tat?aat?gct????1104
Pro?Val?Val?Thr?Glu?Asp?Val?Glu?Gly?Asp?Lys?Lys?Ile?Tyr?Asn?Ala
355?????????????????360?????????????????365
gaa?gtt?gct?gaa?tgg?gat?aag?tta?cgg?caa?aga?ttg?ctt?gat?gcc?aga????1152
Glu?Val?Ala?Glu?Trp?Asp?Lys?Leu?Arg?Gln?Arg?Leu?Leu?Asp?Ala?Arg
370?????????????????375?????????????????380
aat?aaa?atc?acc?tct?gct?gaa?tct?gcg?gta?aat?tcg?gcg?aga?aat?aac????1200
Asn?Lys?Ile?Thr?Ser?Ala?Glu?Ser?Ala?Val?Asn?Ser?Ala?Arg?Asn?Asn
385?????????????????390?????????????????395
ctc?agt?gcc?aga?aca?aat?gag?caa?aag?cat?gca?aat?gac?gct?ctt?aat????1248
Leu?Ser?Ala?Arg?Thr?Asn?Glu?Gln?Lys?His?Ala?Asn?Asp?Ala?Leu?Asn
400?????????????????405?????????????????410?????????????????415
gcc?ctg?ttg?aag?gaa?aaa?gag?aat?ata?cgt?aac?cag?ctt?tcc?ggc?atc????1296
Ala?Leu?Leu?Lys?Glu?Lys?Glu?Asn?Ile?Arg?Asn?Gln?Leu?Ser?Gly?Ile
420?????????????????425?????????????????430
aat?cag?aag?ata?gcg?gaa?gag?aaa?aga?aaa?cag?gat?gaa?ctg?aag?gca????1344
Asn?Gln?Lys?Ile?Ala?Glu?Glu?Lys?Arg?Lys?Gln?Asp?Glu?Leu?Lys?Ala
435?????????????????440?????????????????445
acg?aaa?gac?gca?att?aat?ttc?aca?aca?gag?ttc?ctg?aaa?tca?gtt?tca?????1392
Thr?Lys?Asp?Ala?Ile?Asn?Phe?Thr?Thr?Glu?Phe?Leu?Lys?Ser?Val?Ser
450?????????????????455?????????????????460
gaa?aaa?tat?ggt?gca?aaa?gct?gag?cag?tta?gcc?aga?gag?atg?gcc?ggg?????1440
Glu?Lys?Tyr?Gly?Ala?Lys?Ala?Glu?Gln?Leu?Ala?Arg?Glu?Met?Ala?Gly
465?????????????????470?????????????????475
cag?gct?aaa?ggg?aag?aaa?ata?cgt?aat?gtt?gaa?gag?gca?tta?aaa?acg?????1488
Gln?Ala?Lys?Gly?Lys?Lys?Ile?Arg?Asn?Val?Glu?Glu?Ala?Leu?Lys?Thr
480?????????????????485?????????????????490?????????????????495
tat?gaa?aag?tac?cgg?gct?gac?att?aac?aaa?aaa?att?aat?gca?aaa?gat?????1536
Tyr?Glu?Lys?Tyr?Arg?Ala?Asp?Ile?Asn?Lys?Lys?Ile?Asn?Ala?Lys?Asp
500?????????????????505?????????????????510
cgt?gca?gcg?att?gcc?gca?gcc?ctt?gag?tct?gtg?aag?ctg?tct?gat?ata?????1584
Arg?Ala?Ala?Ile?Ala?Ala?Ala?Leu?Glu?Ser?Val?Lys?Leu?Ser?Asp?Ile
515?????????????????520?????????????????525
tcg?tct?aat?ctg?aac?aga?ttc?agt?cgg?gga?ctg?gga?tat?gca?gga?aaa?????1632
Ser?Ser?Asn?Leu?Asn?Arg?Phe?Ser?Arg?Gly?Leu?Gly?Tyr?Ala?Gly?Lys
530?????????????????535?????????????????540
ttt?aca?agt?ctt?gct?gac?tgg?atc?act?gag?ttt?ggt?aag?gct?gtc?cgg?????1680
Phe?Thr?Ser?Leu?Ala?Asp?Trp?Ile?Thr?Glu?Phe?Gly?Lys?Ala?Val?Arg
545?????????????????550?????????????????555
aca?gag?aac?tgg?cgt?cct?ctt?ttt?gtt?aaa?aca?gaa?acc?atc?ata?gca?????1728
Thr?Glu?Asn?Trp?Arg?Pro?Leu?Phe?Val?Lys?Thr?Glu?Thr?Ile?Ile?Ala
560?????????????????565?????????????????570?????????????????575
ggc?aat?gcc?gca?acg?gct?ctt?gtg?gca?ctg?gtc?ttc?agt?att?ctt?acc?????1776
Gly?Asn?Ala?Ala?Thr?Ala?Leu?Val?Ala?Leu?Val?Phe?Ser?Ile?Leu?Thr
580?????????????????585?????????????????590
gga?agc?gct?tta?ggc?att?atc?ggg?tat?ggt?tta?ctg?atg?gct?gtc?acc?????1824
Gly?Ser?Ala?Leu?Gly?Ile?Ile?Gly?Tyr?Gly?Leu?Leu?Met?Ala?Val?Thr
595?????????????????600?????????????????605
ggt?gcg?ctg?att?gat?gaa?tcg?ctt?gtg?gaa?aaa?gcg?aat?aag?ttc?tgg?????1872
Gly?Ala?Leu?Ile?Asp?Glu?Ser?Leu?Val?Glu?Lys?Ala?Asn?Lys?Phe?Trp
610?????????????????615?????????????????620
ggt?att?gga?gtt?aac?gca?tgt?tct?tcc?ctg?ttt?taa?????????????????????1908
Gly?Ile?Gly?Val?Asn?Ala?Cys?Ser?Ser?Leu?Phe
625?????????????????630
<210>15
<211>634
<212>PRT
＜213〉artificial sequence
<400>15
Ser?Asp?Pro?Val?Arg?Ile?Thr?Asn?Pro?Gly?Ala?Glu?Ser?Leu?Gly?Tyr
1???????????????5???????????????????10??????????????????15
Asp?Ser?Asp?Gly?His?Glu?Ile?Met?Ala?Val?Asp?Ile?Tyr?Val?Asn?Pro
20??????????????????25??????????????????30
Pro?Arg?Val?Asp?Val?Phe?His?Gly?Thr?Pro?Pro?Ala?Trp?Ser?Ser?Phe
35??????????????????40??????????????????45
Gly?Asn?Lys?Thr?Ile?Trp?Gly?Gly?Asn?Glu?Trp?Val?Asp?Asp?Ser?Pro
50??????????????????55??????????????????60
Thr?Arg?Ser?Asp?Ile?Glu?Lys?Arg?Asp?Lys?Glu?Ile?Thr?Ala?Tyr?Lys
65??????????????????70??????????????????75??????????????????80
Asn?Thr?Leu?Ser?Ala?Gln?Gln?Lys?Glu?Asn?Glu?Asn?Lys?Arg?Thr?Glu
85??????????????????90??????????????????95
Ala?Gly?Lys?Arg?Leu?Ser?Ala?Ala?Ile?Ala?Ala?Arg?Glu?Lys?Asp?Glu
100?????????????????105?????????????????110
Asn?Thr?Leu?Lys?Thr?Leu?Arg?Ala?Gly?Asn?Ala?Asp?Ala?Ala?Asp?Ile
115?????????????????120?????????????????125
Thr?Arg?Gln?Glu?Phe?Arg?Leu?Leu?Gln?Ala?Glu?Leu?Arg?Glu?Tyr?Gly
130?????????????????135?????????????????140
Phe?Arg?Thr?Glu?Ile?Ala?Gly?Tyr?Asp?Ala?Leu?Arg?Leu?His?Thr?Glu
145?????????????????150?????????????????155?????????????????160
Ser?Arg?Met?Leu?Phe?Ala?Asp?Ala?Asp?Ser?Leu?Arg?Ile?Ser?Pro?Arg
165?????????????????170?????????????????175
Glu?Ala?Arg?Ser?Leu?Ile?Glu?Gln?Ala?Glu?Lys?Arg?Gln?Lys?Asp?Ala
180?????????????????185?????????????????190
Gln?Asn?Ala?Asp?Lys?Lys?Ala?Ala?Asp?Met?Leu?Ala?Glu?Tyr?Glu?Arg
195?????????????????200?????????????????205
Arg?Lys?Gly?Ile?Leu?Asp?Thr?Arg?Leu?Ser?Glu?Leu?Glu?Lys?Asn?Gly
210?????????????????215?????????????????220
Gly?Ala?Ala?Leu?Ala?Val?Leu?Asp?Ala?Gln?Gln?Ala?Arg?Leu?Leu?Gly
225??????????????????230????????????????235?????????????????240
Glu?Gln?Thr?Arg?Asn?Asp?Arg?Ala?Ile?Ser?Glu?Ala?Arg?Asn?Lys?Leu
245?????????????????250?????????????????255
Ser?Ser?Val?Thr?Glu?Ser?Leu?Asn?Thr?Ala?Arg?Asn?Ala?Leu?Thr?Arg
260?????????????????265?????????????????270
Ala?Glu?Gln?Gln?Leu?Thr?Gln?Gln?Lys?Asn?Thr?Pro?Asp?Gly?Lys?Thr
275?????????????????280?????????????????285
Ile?Val?Ser?Pro?Glu?Lys?Phe?Pro?Gly?Arg?Ser?Ser?Thr?Asn?His?Ser
290?????????????????295?????????????????300
Ile?Val?Val?Ser?Gly?Asp?Pro?Arg?Phe?Ala?Gly?Thr?Ile?Lys?Ile?Thr
305?????????????????310?????????????????315?????????????????320
Thr?Ser?Ala?Val?Ile?Asp?Asn?Arg?Ala?Asn?Leu?Asn?Tyr?Leu?Leu?Ser
325?????????????????330?????????????????335
His?Ser?Gly?Leu?Asp?Tyr?Lys?Arg?Asn?Ile?Leu?Asn?Asp?Arg?Asn?Pro
340?????????????????345?????????????????350
Val?Val?Thr?Glu?Asp?Val?Glu?Gly?Asp?Lys?Lys?Ile?Tyr?Asn?Ala?Glu
355?????????????????360?????????????????365
Val?Ala?Glu?Trp?Asp?Lys?Leu?Arg?Gln?Arg?Leu?Leu?Asp?Ala?Arg?Asn
370?????????????????375?????????????????380
Lys?Ile?Thr?Ser?Ala?Glu?Ser?Ala?Val?Asn?Ser?Ala?Arg?Asn?Asn?Leu
385?????????????????390?????????????????395?????????????????400
Ser?Ala?Arg?Thr?Asn?Glu?Gln?Lys?His?Ala?Asn?Asp?Ala?Leu?Asn?Ala
405?????????????????410?????????????????415
Leu?Leu?Lys?Glu?Lys?Glu?Asn?Ile?Arg?Asn?Gln?Leu?Ser?Gly?Ile?Asn
420?????????????????425?????????????????430
Gln?Lys?Ile?Ala?Glu?Glu?Lys?Arg?Lys?Gln?Asp?Glu?Leu?Lys?Ala?Thr
435?????????????????440?????????????????445
Lys?Asp?Ala?Ile?Asn?Phe?Thr?Thr?Glu?Phe?Leu?Lys?Ser?Val?Ser?Glu
450?????????????????455?????????????????460
Lys?Tyr?Gly?Ala?Lys?Ala?Glu?Gln?Leu?Ala?Arg?Glu?Met?Ala?Gly?Gln
465?????????????????470?????????????????475?????????????????480
Ala?Lys?Gly?Lys?Lys?Ile?Arg?Asn?Val?Glu?Glu?Ala?Leu?Lys?Thr?Tyr
485?????????????????490?????????????????495
Glu?Lys?Tyr?Arg?Ala?Asp?Ile?Asn?Lys?Lys?Ile?Asn?Ala?Lys?Asp?Arg
500?????????????????505?????????????????510
Ala?Ala?Ile?Ala?Ala?Ala?Leu?Glu?Ser?Val?Lys?Leu?Ser?Asp?Ile?Ser
515?????????????????520?????????????????525
Ser?Asn?Leu?Asn?Arg?Phe?Ser?Arg?Gly?Leu?Gly?Tyr?Ala?Gly?Lys?Phe
530?????????????????535?????????????????540
Thr?Ser?Leu?Ala?Asp?Trp?Ile?Thr?Glu?Phe?Gly?Lys?Ala?Val?Arg?Thr
545?????????????????550?????????????????555?????????????????560
Glu?Asn?Trp?Arg?Pro?Leu?Phe?Val?Lys?Thr?Glu?Thr?Ile?Ile?Ala?Gly
565?????????????????570?????????????????575
Asn?Ala?Ala?Thr?Ala?Leu?Val?Ala?Leu?Val?Phe?Ser?Ile?Leu?Thr?Gly
580?????????????????585?????????????????590
Ser?Ala?Leu?Gly?Ile?Ile?Gly?Tyr?Gly?Leu?Leu?Met?Ala?Val?Thr?Gly
595?????????????????600?????????????????605
Ala?Leu?Ile?Asp?Glu?Ser?Leu?Val?Glu?Lys?Ala?Asn?Lys?Phe?Trp?Gly
610?????????????????615?????????????620
Ile?Gly?Val?Asn?Ala?Cys?Ser?Ser?Leu?Phe
625?????????????????630
<210>16
<211>1905
<212>DNA
＜213〉artificial sequence
<220>
<221>CDS
<222>(4)..(1902)
＜223〉coding staphylococcus polypeptide
<400>16
atg?tct?gac?cct?gta?cgt?att?aca?aat?ccc?ggt?gca?gaa?tcg?ctg?ggg???????48
Ser?Asp?Pro?Val?Arg?Ile?Thr?Asn?Pro?Gly?Ala?Glu?Ser?Leu?Gly
1???????????????5???????????????????10??????????????????15
tat?gat?tca?gat?ggc?cat?gaa?att?atg?gcc?gtt?gat?att?tat?gta?aac?????96
Tyr?Asp?Ser?Asp?Gly?His?Glu?Ile?Met?Ala?Val?Asp?Ile?Tyr?Val?Asn
20??????????????????25??????????????????30
cct?cca?cgt?gtc?gat?gtc?ttt?cat?ggt?acc?ccg?cct?gca?tgg?agt?tcc????144
Pro?Pro?Arg?Val?Asp?Val?Phe?His?Gly?Thr?Pro?Pro?Ala?Trp?Ser?Ser
35??????????????????40??????????????????45
ttc?ggg?aac?aaa?acc?atc?tgg?ggc?gga?aac?gag?tgg?gtt?gat?gat?tcc????192
Phe?Gly?Asn?Lys?Thr?Ile?Trp?Gly?Gly?Asn?Glu?Trp?Val?Asp?Asp?Ser
50??????????????????55??????????????????60
cca?acc?cga?agt?gat?atc?gaa?aaa?agg?gac?aag?gaa?atc?aca?gcg?tac????240
Pro?Thr?Arg?Ser?Asp?Ile?Glu?Lys?Arg?Asp?Lys?Glu?Ile?Thr?Ala?Tyr
65??????????????????70??????????????????75
aaa?aac?acg?ctc?agc?gcg?cag?cag?aaa?gag?aat?gag?aat?aag?cgt?act????288
Lys?Asn?Thr?Leu?Ser?Ala?Gln?Gln?Lys?Glu?Asn?Glu?Asn?Lys?Arg?Thr
80??????????????????85??????????????????90??????????????????95
gaa?gcc?gga?aaa?cgc?ctc?tct?gcg?gcg?att?gct?gca?agg?gaa?aaa?gat????336
Glu?Ala?Gly?Lys?Arg?Leu?Ser?Ala?Ala?Ile?Ala?Ala?Arg?Glu?Lys?Asp
100?????????????????105?????????????????110
gaa?aac?aca?ctg?aaa?aca?ctc?cgt?gcc?gga?aac?gca?gat?gcc?gct?gat????384
Glu?Asn?Thr?Leu?Lys?Thr?Leu?Arg?Ala?Gly?Asn?Ala?Asp?Ala?Ala?Asp
115?????????????????120?????????????????125
att?aca?cga?cag?gag?ttc?aga?ctc?ctg?cag?gca?gag?ctg?aga?gaa?tac????432
Ile?Thr?Arg?Gln?Glu?Phe?Arg?Leu?Leu?Gln?Ala?Glu?Leu?Arg?Glu?Tyr
130?????????????????135?????????????????140
gga?ttc?cgt?act?gaa?atc?gcc?gga?tat?gac?gcc?ctc?cgg?ctg?cat?aca????480
Gly?Phe?Arg?Thr?Glu?Ile?Ala?Gly?Tyr?Asp?Ala?Leu?Arg?Leu?His?Thr
145?????????????????150?????????????????155
gag?agc?cgg?atg?ctg?ttt?gct?gat?gct?gat?tct?ctt?cgt?ata?tct?ccc????528
Glu?Ser?Arg?Met?Leu?Phe?Ala?Asp?Ala?Asp?Ser?Leu?Arg?Ile?Ser?Pro
160?????????????????165?????????????????170?????????????????175
cgg?gag?gcc?agg?tcg?tta?atc?gaa?cag?gct?gaa?aaa?cgg?cag?aag?gat????576
Arg?Glu?Ala?Arg?Ser?Leu?Ile?Glu?Gln?Ala?Glu?Lys?Arg?Gln?Lys?Asp
180?????????????????185?????????????????190
gcg?cag?aac?gca?gac?aag?aag?gcc?gct?gat?atg?ctt?gct?gaa?tac?gag????624
Ala?Gln?Asn?Ala?Asp?Lys?Lys?Ala?Ala?Asp?Met?Leu?Ala?Glu?Tyr?Glu
195?????????????????200?????????????????205
cgc?aga?aaa?ggt?att?ctg?gac?acc?cgg?ttg?tca?gag?ctg?gaa?aaa?aat????672
Arg?Arg?Lys?Gly?Ile?Leu?Asp?Thr?Arg?Leu?Ser?Glu?Leu?Glu?Lys?Asn
210?????????????????215?????????????????220
ggc?ggg?gca?gcc?ctt?gcc?gtt?ctt?gat?gca?caa?cag?gcc?cgt?ctg?ctc????720
Gly?Gly?Ala?Ala?Leu?Ala?Val?Leu?Asp?Ala?Gln?Gln?Ala?Arg?Leu?Leu
225?????????????????230?????????????????235
ggg?cag?cag?aca?cgg?aat?gac?agg?gcc?att?tca?gag?gcc?cgg?aat?aaa????768
Gly?Gln?Gln?Thr?Arg?Asn?Asp?Arg?Ala?Ile?Ser?Glu?Ala?Arg?Asn?Lys
240?????????????????245?????????????????250?????????????????255
ctc?agt?tca?gtg?acg?gaa?tcg?ctt?aac?acg?gcc?cgt?aat?gca?tta?acc????816
Leu?Ser?Ser?Val?Thr?Glu?Ser?Leu?Asn?Thr?Ala?Arg?Asn?Ala?Leu?Thr
260?????????????????265?????????????????270
aga?gct?gaa?caa?cag?ctg?acg?caa?cag?aaa?aac?acg?cct?gac?ggc?aaa????864
Arg?Ala?Glu?Gln?Gln?Leu?Thr?Gln?Gln?Lys?Asn?Thr?Pro?Asp?Gly?Lys
275?????????????????280?????????????????285
acg?ata?gtt?tcc?cct?gaa?aaa?ttc?ccg?ggg?cgt?tca?tca?aca?aat?cat????912
Thr?Ile?Val?Ser?Pro?Glu?Lys?Phe?Pro?Gly?Arg?Ser?Ser?Thr?Asn?His
290?????????????????295?????????????????300
tct?att?gtt?gtg?agc?ggt?gat?ccg?aga?ttt?gcc?ggt?acg?ata?aaa?atc????960
Ser?Ile?Val?Val?Ser?Gly?Asp?Pro?Arg?Phe?Ala?Gly?Thr?Ile?Lys?Ile
305?????????????????310?????????????????315
aca?acc?agc?gca?gtc?atc?gat?aac?cgt?gca?aac?ctg?aat?tat?ctt?ctg????1008
Thr?Thr?Ser?Ala?Val?Ile?Asp?Asn?Arg?Ala?Asn?Leu?Asn?Tyr?Leu?Leu
320?????????????????325?????????????????330?????????????????335
agc?cat?tcc?ggt?ctg?gac?tat?aaa?cgc?aat?att?ctg?aat?gac?cgg?aat????1056
Ser?His?Ser?Gly?Leu?Asp?Tyr?Lys?Arg?Asn?Ile?Leu?Asn?Asp?Arg?Asn
340?????????????????345?????????????????350
ccg?gtg?gtg?aca?gag?gat?gtg?gaa?ggt?gac?aag?aaa?att?tat?aat?gct????1104
Pro?Val?Val?Thr?Glu?Asp?Val?Glu?Gly?Asp?Lys?Lys?Ile?Tyr?Asn?Ala
355?????????????????360?????????????????365
gaa?gtt?gct?gaa?tgg?gat?aag?tta?cgg?caa?aga?ttg?ctt?gat?gcc?aga????1152
Glu?Val?Ala?Glu?Trp?Asp?Lys?Leu?Arg?Gln?Arg?Leu?Leu?Asp?Ala?Arg
370?????????????????375?????????????????380
aat?aaa?atc?acc?tct?gct?gaa?tct?gcg?gta?aat?tcg?gcg?aga?aat?aac????1200
Asn?Lys?Ile?Thr?Ser?Ala?Glu?Ser?Ala?Val?Asn?Ser?Ala?Arg?Asn?Asn
385?????????????????390?????????????????395
ctc?agt?gcc?aga?aca?aat?gag?caa?aag?cat?gca?aat?gac?gct?ctt?aat????1248
Leu?Ser?Ala?Arg?Thr?Asn?Glu?Gln?Lys?His?Ala?Asn?Asp?Ala?Leu?Asn
400?????????????????405?????????????????410?????????????????415
gcc?ctg?ttg?aag?gaa?aaa?gag?aat?ata?cgt?aac?cag?ctt?tcc?ggc?atc????1296
Ala?Leu?Leu?Lys?Glu?Lys?Glu?Asn?Ile?Arg?Asn?Gln?Leu?Ser?Gly?Ile
420?????????????????425?????????????????430
aat?cag?aag?ata?gcg?gaa?gag?aaa?aga?aaa?cag?gat?gaa?ctg?aag?gca????1344
Asn?Gln?Lys?Ile?Ala?Glu?Glu?Lys?Arg?Lys?Gln?Asp?Glu?Leu?Lys?Ala
435?????????????????440?????????????????445
acg?aaa?gac?gca?att?aat?ttc?aca?aca?gag?ttc?ctg?aaa?tca?gtt?tca????1392
Thr?Lys?Asp?Ala?Ile?Asn?Phe?Thr?Thr?Glu?Phe?Leu?Lys?Ser?Val?Ser
450?????????????????455?????????????????460
gaa?aaa?tat?ggt?gca?aaa?gct?gag?cag?tta?gcc?aga?gag?atg?gcc?ggg????1440
Glu?Lys?Tyr?Gly?Ala?Lys?Ala?Glu?Gln?Leu?Ala?Arg?Glu?Met?Ala?Gly
465?????????????????470?????????????????475
cag?gct?aaa?ggg?aag?aaa?ata?cgt?aat?gtt?gaa?gag?gca?tta?aaa?acg????1488
Gln?Ala?Lys?Gly?Lys?Lys?Ile?Arg?Asn?Val?Glu?Glu?Ala?Leu?Lys?Thr
480?????????????????485?????????????????490?????????????????495
tat?gaa?aag?tac?cgg?gct?gac?att?aac?aaa?aaa?att?aat?gca?aaa?gat????1536
Tyr?Glu?Lys?Tyr?Arg?Ala?Asp?Ile?Asn?Lys?Lys?Ile?Asn?Ala?Lys?Asp
500?????????????????505?????????????????510
cgt?gca?gcg?att?gcc?gca?gcc?ctt?gag?tct?gtg?aag?ctg?tct?gat?ata????1584
Arg?Ala?Ala?Ile?Ala?Ala?Ala?Leu?Glu?Ser?Val?Lys?Leu?Ser?Asp?Ile
515?????????????????520?????????????????525
tcg?tct?aat?ctg?aac?aga?ttc?agt?cgg?gga?ctg?gga?tat?gca?gga?aaa????1632
Ser?Ser?Asn?Leu?Asn?Arg?Phe?Ser?Arg?Gly?Leu?Gly?Tyr?Ala?Gly?Lys
530?????????????????535?????????????????540
ttt?aca?agt?ctt?gct?gac?tgg?atc?act?gag?ttt?ggt?aag?gct?gtc?cgg????1680
Phe?Thr?Ser?Leu?Ala?Asp?Trp?Ile?Thr?Glu?Phe?Gly?Lys?Ala?Val?Arg
545?????????????????550?????????????????555
aca?gag?aac?tgg?cgt?cct?ctt?ttt?gtt?aaa?aca?gaa?acc?atc?ata?gca????1728
Thr?Glu?Asn?Trp?Arg?Pro?Leu?Phe?Val?Lys?Thr?Glu?Thr?Ile?Ile?Ala
560?????????????????565?????????????????570?????????????????575
ggc?aat?gcc?gca?acg?gct?ctt?gtg?gca?ctg?gtc?ttc?agt?att?ctt?acc????1776
Gly?Asn?Ala?Ala?Thr?Ala?Leu?Val?Ala?Leu?Val?Phe?Ser?Ile?Leu?Thr
580?????????????????585?????????????????590
gga?agc?gct?tta?ggc?att?atc?ggg?tat?ggt?tta?ctg?atg?gct?gtc?acc????1824
Gly?Ser?Ala?Leu?Gly?Ile?Ile?Gly?Tyr?Gly?Leu?Leu?Met?Ala?Val?Thr
595?????????????????600?????????????????605
ggt?gcg?ctg?att?gat?gaa?tcg?ctt?gtg?gaa?aaa?gcg?aat?aag?ttc?tgg????1872
Gly?Ala?Leu?Ile?Asp?Glu?Ser?Leu?Val?Glu?Lys?Ala?Asn?Lys?Phe?Trp
610?????????????????????615?????????????????620
ggt?att?tat?ata?aac?tgt?gat?ttt?ctt?ctg?taa????????????????????????1905
Gly?Ile?Tyr?Ile?Asn?Cys?Asp?Phe?Leu?Leu
625?????????????????630
<210>17
<211>633
<212>PRT
＜213〉artificial sequence
<400>17
Ser?Asp?Pro?Val?Arg?Ile?Thr?Asn?Pro?Gly?Ala?Glu?Ser?Leu?Gly?Tyr
1???????????????5???????????????????10??????????????????15
Asp?Ser?Asp?Gly?His?Glu?Ile?Met?Ala?Val?Asp?Ile?Tyr?Val?Asn?Pro
20??????????????????25??????????????????30
Pro?Arg?Val?Asp?Val?Phe?His?Gly?Thr?Pro?Pro?Ala?Trp?Ser?Ser?Phe
35??????????????????40??????????????????45
Gly?Asn?Lys?Thr?Ile?Trp?Gly?Gly?Asn?Glu?Trp?Val?Asp?Asp?Ser?Pro
50??????????????????55??????????????????60
Thr?Arg?Ser?Asp?Ile?Glu?Lys?Arg?Asp?Lys?Glu?Ile?Thr?Ala?Tyr?Lys
65??????????????????70??????????????????75??????????????????80
Asn?Thr?Leu?Ser?Ala?Gln?Gln?Lys?Glu?Asn?Glu?Asn?Lys?Arg?Thr?Glu
85??????????????????90??????????????????95
Ala?Gly?Lys?Arg?Leu?Ser?Ala?Ala?Ile?Ala?Ala?Arg?Glu?Lys?Asp?Glu
100?????????????????105?????????????????110
Asn?Thr?Leu?Lys?Thr?Leu?Arg?Ala?Gly?Asn?Ala?Asp?Ala?Ala?Asp?Ile
115?????????????????120?????????????????125
Thr?Arg?Gln?Glu?Phe?Arg?Leu?Leu?Gln?Ala?Glu?Leu?Arg?Glu?Tyr?Gly
130?????????????????135?????????????????140
Phe?Arg?Thr?Glu?Ile?Ala?Gly?Tyr?Asp?Ala?Leu?Arg?Leu?His?Thr?Glu
145?????????????????150?????????????????155?????????????????160
Ser?Arg?Met?Leu?Phe?Ala?Asp?Ala?Asp?Ser?Leu?Arg?Ile?Ser?Pro?Arg
165?????????????????170?????????????????175
Glu?Ala?Arg?Ser?Leu?Ile?Glu?Gln?Ala?Glu?Lys?Arg?Gln?Lys?Asp?Ala
180?????????????????185?????????????????190
Gln?Asn?Ala?Asp?Lys?Lys?Ala?Ala?Asp?Met?Leu?Ala?Glu?Tyr?Glu?Arg
195?????????????????200?????????????????205
Arg?Lys?Gly?Ile?Leu?Asp?Thr?Arg?Leu?Ser?Glu?Leu?Glu?Lys?Asn?Gly
210?????????????????215?????????????????220
Gly?Ala?Ala?Leu?Ala?Val?Leu?Asp?Ala?Gln?Gln?Ala?Arg?Leu?Leu?Gly
225?????????????????230?????????????????235?????????????????240
Gln?Gln?Thr?Arg?Asn?Asp?Arg?Ala?Ile?Ser?Glu?Ala?Arg?Asn?Lys?Leu
245?????????????????250?????????????????255
Ser?Ser?Val?Thr?Glu?Ser?Leu?Asn?Thr?Ala?Arg?Asn?Ala?Leu?Thr?Arg
260?????????????????265?????????????????270
Ala?Glu?Gln?Gln?Leu?Thr?Gln?Gln?Lys?Asn?Thr?Pro?Asp?Gly?Lys?Thr
275?????????????????280?????????????????285
Ile?Val?Ser?Pro?Glu?Lys?Phe?Pro?Gly?Arg?Ser?Ser?Thr?Asn?His?Ser
290?????????????????295?????????????????300
Ile?Val?Val?Ser?Gly?Asp?Pro?Arg?Phe?Ala?Gly?Thr?Ile?Lys?Ile?Thr
305?????????????????310?????????????????315?????????????????320
Thr?Ser?Ala?Val?Ile?Asp?Asn?Arg?Ala?Asn?Leu?Asn?Tyr?Leu?Leu?Ser
325?????????????????330?????????????????335
His?Ser?Gly?Leu?Asp?Tyr?Lys?Arg?Asn?Ile?Leu?Asn?Asp?Arg?Asn?Pro
340?????????????????345?????????????????350
Val?Val?Thr?Glu?Asp?Val?Glu?Gly?Asp?Lys?Lys?Ile?Tyr?Asn?Ala?Glu
355?????????????????360?????????????????365
Val?Ala?Glu?Trp?Asp?Lys?Leu?Arg?Gln?Arg?Leu?Leu?Asp?Ala?Arg?Asn
370?????????????????375?????????????????380
Lys?Ile?Thr?Ser?Ala?Glu?Ser?Ala?Val?Asn?Ser?Ala?Arg?Asn?Asn?Leu
385?????????????????390?????????????????395?????????????????400
Ser?Ala?Arg?Thr?Asn?Glu?Gln?Lys?His?Ala?Asn?Asp?Ala?Leu?Asn?Ala
405?????????????????410?????????????????415
Leu?Leu?Lys?Glu?Lys?Glu?Asn?Ile?Arg?Asn?Gln?Leu?Ser?Gly?Ile?Asn
420?????????????????425?????????????????430
Gln?Lys?Ile?Ala?Glu?Glu?Lys?Arg?Lys?Gln?Asp?Glu?Leu?Lys?Ala?Thr
435?????????????????440?????????????????445
Lys?Asp?Ala?Ile?Asn?Phe?Thr?Thr?Glu?Phe?Leu?Lys?Ser?Val?Ser?Glu
450?????????????????455?????????????????460
Lys?Tyr?Gly?Ala?Lys?Ala?Glu?Gln?Leu?Ala?Arg?Glu?Met?Ala?Gly?Gln
465?????????????????470?????????????????475?????????????????480
Ala?Lys?Gly?Lys?Lys?Ile?Arg?Asn?Val?Glu?Glu?Ala?Leu?Lys?Thr?Tyr
485?????????????????490?????????????????495
Glu?Lys?Tyr?Arg?Ala?Asp?Ile?Asn?Lys?Lys?Ile?Asn?Ala?Lys?Asp?Arg
500??????????????????505?????????????????510
Ala?Ala?Ile?Ala?Ala?Ala?Leu?Glu?Ser?Val?Lys?Leu?Ser?Asp?Ile?Ser
515?????????????????520????????????????525
Ser?Asn?Leu?Asn?Arg?Phe?Ser?Arg?Gly?Leu?Gly?Tyr?Ala?Gly?Lys?Phe
530?????????????????535?????????????????540
Thr?Ser?Leu?Ala?Asp?Trp?Ile?Thr?Glu?Phe?Gly?Lys?Ala?Val?Arg?Thr
545?????????????????550?????????????????555?????????????????560
Glu?Asn?Trp?Arg?Pro?Leu?Phe?Val?Lys?Thr?Glu?Thr?Ile?Ile?Ala?Gly
565?????????????????570?????????????????575
Asn?Ala?Ala?Thr?Ala?Leu?Val?Ala?Leu?Val?Phe?Ser?Ile?Leu?Thr?Gly
580?????????????????585?????????????????590
Ser?Ala?Leu?Gly?Ile?Ile?Gly?Tyr?Gly?Leu?Leu?Met?Ala?Val?Thr?Gly
595?????????????????600?????????????????605
Ala?Leu?Ile?Asp?Glu?Ser?Leu?Val?Glu?Lys?Ala?Asn?Lys?Phe?Trp?Gly
610?????????????????615?????????????????620
Ile?Tyr?Ile?Asn?Cys?Asp?Phe?Leu?Leu
625?????????????????630
<210>18
<211>1905
<212>DNA
＜213〉artificial sequence
<220>
<221>CDS
<222>(4)..(1902)
＜223〉coding staphylococcus polypeptide
<400>18
atg?tct?gac?cct?gta?cgt?att?aca?aat?ccc?ggt?gca?gaa?tcg?ctg?ggg?????48
Ser?Asp?Pro?Val?Arg?Ile?Thr?Asn?Pro?Gly?Ala?Glu?Ser?Leu?Gly
1???????????????5???????????????????10??????????????????15
tat?gat?tca?gat?ggc?cat?gaa?att?atg?gcc?gtt?gat?att?tat?gta?aac?????96
Tyr?Asp?Ser?Asp?Gly?His?Glu?Ile?Met?Ala?Val?Asp?Ile?Tyr?Val?Asn
20??????????????????25??????????????????30
cct?cca?cgt?gtc?gat?gtc?ttt?cat?ggt?acc?ccg?cct?gca?tgg?agt?tcc????144
Pro?Pro?Arg?Val?Asp?Val?Phe?His?Gly?Thr?Pro?Pro?Ala?Trp?Ser?Ser
35??????????????????40??????????????????45
ttc?ggg?aac?aaa?acc?atc?tgg?ggc?gga?aac?gag?tgg?gtt?gat?gat?tcc????192
Phe?Gly?Asn?Lys?Thr?Ile?Trp?Gly?Gly?Asn?Glu?Trp?Val?Asp?Asp?Ser
50??????????????????55??????????????????60
cca?acc?cga?agt?gat?atc?gaa?aaa?agg?gac?aag?gaa?atc?aca?gcg?tac????240
Pro?Thr?Arg?Ser?Asp?Ile?Glu?Lys?Arg?Asp?Lys?Glu?Ile?Thr?Ala?Tyr
65??????????????????70??????????????????75
aaa?aac?acg?ctc?agc?gcg?cag?cag?aaa?gag?aat?gag?aat?aag?cgt?act????288
Lys?Asn?Thr?Leu?Ser?Ala?Gln?Gln?Lys?Glu?Asn?Glu?Asn?Lys?Arg?Thr
80??????????????????85??????????????????90??????????????????95
gaa?gcc?gga?aaa?cgc?ctc?tct?gcg?gcg?att?gct?gca?agg?gaa?aaa?gat????336
Glu?Ala?Gly?Lys?Arg?Leu?Ser?Ala?Ala?Ile?Ala?Ala?Arg?Glu?Lys?Asp
100?????????????????105?????????????????110
gaa?aac?aca?ctg?aaa?aca?ctc?cgt?gcc?gga?aac?gca?gat?gcc?gct?gat????384
Glu?Asn?Thr?Leu?Lys?Thr?Leu?Arg?Ala?Gly?Asn?Ala?Asp?Ala?Ala?Asp
115?????????????????120?????????????????125
att?aca?cga?cag?gag?ttc?aga?ctc?ctg?cag?gca?gag?ctg?aga?gaa?tac????432
Ile?Thr?Arg?Gln?Glu?Phe?Arg?Leu?Leu?Gln?Ala?Glu?Leu?Arg?Glu?Tyr
130?????????????????135?????????????????140
gga?ttc?cgt?act?gaa?atc?gcc?gga?tat?gac?gcc?ctc?cgg?ctg?cat?aca????480
Gly?Phe?Arg?Thr?Glu?Ile?Ala?Gly?Tyr?Asp?Ala?Leu?Arg?Leu?His?Thr
145?????????????????150?????????????????155
gag?agc?cgg?atg?ctg?ttt?gct?gat?gct?gat?tct?ctt?cgt?ata?tct?ccc????528
Glu?Ser?Arg?Met?Leu?Phe?Ala?Asp?Ala?Asp?Ser?Leu?Arg?Ile?Ser?Pro
160?????????????????165??????????????????170????????????????175
cgg?gag?gcc?agg?tcg?tta?atc?gaa?cag?gct?gaa?aaa?cgg?cag?aag?gat????576
Arg?Glu?Ala?Arg?Ser?Leu?Ile?Glu?Gln?Ala?Glu?Lys?Arg?Gln?Lys?Asp
180?????????????????185?????????????????190
gcg?cag?aac?gca?gac?aag?aag?gcc?gct?gat?atg?ctt?gct?gaa?tac?gag??????624
Ala?Gln?Asn?Ala?Asp?Lys?Lys?Ala?Ala?Asp?Met?Leu?Ala?Glu?Tyr?Glu
195?????????????????200?????????????????205
cgc?aga?aaa?ggt?att?ctg?gac?acc?cgg?ttg?tca?gag?ctg?gaa?aaa?aat??????672
Arg?Arg?Lys?Gly?Ile?Leu?Asp?Thr?Arg?Leu?Ser?Glu?Leu?Glu?Lys?Asn
210?????????????????215?????????????????220
ggc?ggg?gca?gcc?ctt?gcc?gtt?ctt?gat?gca?caa?cag?gcc?cgt?ctg?ctc??????720
Gly?Gly?Ala?Ala?Leu?Ala?Val?Leu?Asp?Ala?Gln?Gln?Ala?Arg?Leu?Leu
225?????????????????230?????????????????235
ggg?cag?cag?aca?cgg?aat?gac?agg?gcc?att?tca?gag?gcc?cgg?aat?aaa??????768
Gly?Gln?Gln?Thr?Arg?Asn?Asp?Arg?Ala?Ile?Ser?Glu?Ala?Arg?Asn?Lys
240?????????????????245?????????????????250?????????????????255
ctc?agt?tca?gtg?acg?gaa?tcg?crt?aac?acg?gcc?cgt?aat?gca?tta?acc??????816
Leu?Ser?Ser?Val?Thr?Glu?Ser?Leu?Asn?Thr?Ala?Arg?Asn?Ala?Leu?Thr
260?????????????????265?????????????????270
aga?gct?gaa?caa?cag?ctg?acg?caa?cag?aaa?aac?acg?cct?gac?ggc?aaa??????864
Arg?Ala?Glu?Gln?Gln?Leu?Thr?Gln?Gln?Lys?Asn?Thr?Pro?Asp?Gly?Lys
275?????????????????280?????????????????285
acg?ata?gtt?tcc?cct?gaa?aaa?ttc?ccg?ggg?cgt?tca?tca?aca?aat?cat??????912
Thr?Ile?Val?Ser?Pro?Glu?Lys?Phe?Pro?Gly?Arg?Ser?Ser?Thr?Asn?His
290?????????????????295?????????????????300
tct?att?gtt?gtg?agc?ggt?gat?ccg?aga?ttt?gcc?ggt?acg?ata?aaa?atc??????960
Ser?Ile?Val?Val?Ser?Gly?Asp?Pro?Arg?Phe?Ala?Gly?Thr?Ile?Lys?Ile
305?????????????????310?????????????????315
aca?acc?agc?gca?gtc?atc?gat?aac?cgt?gca?aac?ctg?aat?tat?ctt?ctg?????1008
Thr?Thr?Ser?Ala?Val?Ile?Asp?Asn?Arg?Ala?Asn?Leu?Asn?Tyr?Leu?Leu
320?????????????????325?????????????????330?????????????????335
agc?cat?tcc?ggt?ctg?gac?tat?aaa?cgc?aat?att?ctg?aat?gac?cgg?aat?????1056
Ser?His?Ser?Gly?Leu?Asp?Tyr?Lys?Arg?Asn?Ile?Leu?Asn?Asp?Arg?Asn
340?????????????????345?????????????????350
ccg?gtg?gtg?aca?gag?gat?gtg?gaa?ggt?gac?aag?aaa?att?tat?aat?gct?????1104
Pro?Val?Val?Thr?Glu?Asp?Val?Glu?Gly?Asp?Lys?Lys?Ile?Tyr?Asn?Ala
355?????????????????360?????????????????365
gaa?gtt?gct?gaa?tgg?gat?aag?tta?cgg?caa?aga?ttg?ctt?gat?gcc?aga?????1152
Glu?Val?Ala?Glu?Trp?Asp?Lys?Leu?Arg?Gln?Arg?Leu?Leu?Asp?Ala?Arg
370?????????????????375?????????????????380
aat?aaa?atc?acc?tct?gct?gaa?tct?gcg?gta?aat?tcg?gcg?aga?aat?aac?????1200
Asn?Lys?Ile?Thr?Ser?Ala?Glu?Ser?Ala?Val?Asn?Ser?Ala?Arg?Asn?Asn
385?????????????????390?????????????????395
ctc?agt?gcc?aga?aca?aat?gag?caa?aag?cat?gca?aat?gac?gct?ctt?aat?????1248
Leu?Ser?Ala?Arg?Thr?Asn?Glu?Gln?Lys?His?Ala?Asn?Asp?Ala?Leu?Asn
400?????????????????405?????????????????410?????????????????415
gcc?ctg?ttg?aag?gaa?aaa?gag?aat?ata?cgt?aac?cag?ctt?tcc?ggc?atc?????1296
Ala?Leu?Leu?Lys?Glu?Lys?Glu?Asn?Ile?Arg?Asn?Gln?Leu?Ser?Gly?Ile
420?????????????????425?????????????????430
aat?cag?aag?ata?gcg?gaa?gag?aaa?aga?aaa?cag?gat?gaa?ctg?aag?gca?????1344
Asn?Gln?Lys?Ile?Ala?Glu?Glu?Lys?Arg?Lys?Gln?Asp?Glu?Leu?Lys?Ala
435?????????????????440?????????????????445
acg?aaa?gac?gca?att?aat?ttc?aca?aca?gag?ttc?ctg?aaa?tca?gtt?tca?????1392
Thr?Lys?Asp?Ala?Ile?Asn?Phe?Thr?Thr?Glu?Phe?Leu?Lys?Ser?Val?Ser
450?????????????????455?????????????????460
gaa?aaa?tat?ggt?gca?aaa?gct?gag?cag?tta?gcc?aga?gag?atg?gcc?ggg?????1440
Glu?Lys?Tyr?Gly?Ala?Lys?Ala?Glu?Gln?Leu?Ala?Arg?Glu?Met?Ala?Gly
465?????????????????470?????????????????475
cag?gct?aaa?ggg?aag?aaa?ata?cgt?aat?gtt?gaa?gag?gca?tta?aaa?acg?????1488
Gln?Ala?Lys?Gly?Lys?Lys?Ile?Arg?asn?Val?Glu?Glu?ala?Leu?Lys?Thr
480?????????????????485?????????????????490?????????????????495
tat?gaa?aag?tac?cgg?gct?gac?att?aac?aaa?aaa?att?aat?gca?aaa?gat?????1536
Tyr?Glu?Lys?Tyr?Arg?Ala?Asp?Ile?Asn?Lys?Lys?Ile?Asn?Ala?Lys?Asp
500?????????????????505?????????????????510
cgt?gca?gcg?att?gcc?gca?gcc?ctt?gag?tct?gtg?aag?ctg?tct?gat?ata????1584
Arg?Ala?Ala?Ile?Ala?Ala?Ala?Leu?Glu?Ser?Val?Lys?Leu?Ser?Asp?Ile
515?????????????????520?????????????????525
tcg?tct?aat?ctg?aac?aga?ttc?agt?cgg?gga?ctg?gga?tat?gca?gga?aaa????1632
Ser?Ser?Asn?Leu?Asn?Arg?Phe?Ser?Arg?Gly?Leu?Gly?Tyr?Ala?Gly?Lys
530?????????????????535?????????????????540
ttt?aca?agt?ctt?gct?gac?tgg?atc?act?gag?ttt?ggt?aag?gct?gtc?cgg????1680
Phe?Thr?Ser?Leu?Ala?Asp?Trp?Ile?Thr?Glu?Phe?Gly?Lys?Ala?Val?Arg
545?????????????????550?????????????????555
aca?gag?aac?tgg?cgt?cct?ctt?ttt?gtt?aaa?aca?gaa?acc?atc?ata?gca????1728
Thr?Glu?Asn?Trp?Arg?Pro?Leu?Phe?Val?Lys?Thr?Glu?Thr?Ile?Ile?Ala
560?????????????????565?????????????????570?????????????????575
ggc?aat?gcc?gca?acg?gct?ctt?gtg?gca?ctg?gtc?ttc?agt?att?ctt?acc????1776
Gly?Asn?Ala?Ala?Thr?Ala?Leu?Val?Ala?Leu?Val?Phe?Ser?Ile?Leu?Thr
580?????????????????585?????????????????590
gga?agc?gct?tta?ggc?att?atc?ggg?tat?ggt?tta?ctg?atg?gct?gtc?acc????1824
Gly?Ser?Ala?Leu?Gly?Ile?Ile?Gly?Tyr?Gly?Leu?Leu?Met?Ala?Val?Thr
595?????????????????600?????????????????605
ggt?gcg?ctg?att?gat?gaa?tcg?ctt?gtg?gaa?aaa?gcg?aat?aag?ttc?tgg????1872
Gly?Ala?Leu?Ile?Asp?Glu?Ser?Leu?Val?Glu?Lys?Ala?Asn?Lys?Phe?Trp
610?????????????????615?????????????????620
ggt?att?tat?tcc?acc?tgt?tac?ttt?ata?atg?taa????????????????????????1905
Gly?Ile?Tyr?Ser?Thr?Cys?Tyr?Phe?Ile?Met
625?????????????????630
<210>19
<211>633
<212>PRT
＜213〉artificial sequence
<400>19
Ser?Asp?Pro?Val?Arg?Ile?Thr?Asn?Pro?Gly?Ala?Glu?Ser?Leu?Gly?Tyr
1???????????????5???????????????????10??????????????????15
Asp?Ser?Asp?Gly?His?Glu?Ile?Met?Ala?Val?Asp?Ile?Tyr?Val?Asn?Pro
20??????????????????25??????????????????30
Pro?Arg?Val?Asp?Val?Phe?His?Gly?Thr?Pro?Pro?Ala?Trp?Ser?Ser?Phe
35??????????????????40??????????????????45
Gly?Asn?Lys?Thr?Ile?Trp?Gly?Gly?Asn?Glu?Trp?Val?Asp?Asp?Ser?Pro
50??????????????????55??????????????????60
Thr?Arg?Ser?Asp?Ile?Glu?Lys?Arg?Asp?Lys?Glu?Ile?Thr?Ala?Tyr?Lys
65??????????????????70??????????????????75??????????????????80
Asn?Thr?Leu?Ser?Ala?Gln?Gln?Lys?Glu?Asn?Glu?Asn?Lys?Arg?Thr?Glu
85??????????????????90??????????????????95
Ala?Gly?Lys?Arg?Leu?Ser?Ala?Ala?Ile?Ala?Ala?Arg?Glu?Lys?Asp?Glu
100?????????????????105?????????????????110
Asn?Thr?Leu?Lys?Thr?Leu?Arg?Ala?Gly?Asn?Ala?Asp?Ala?Ala?Asp?Ile
115?????????????????120?????????????????125
Thr?Arg?Gln?Glu?Phe?Arg?Leu?Leu?Gln?Ala?Glu?Leu?Arg?Glu?Tyr?Gly
130?????????????????135?????????????????140
Phe?Arg?Thr?Glu?Ile?Ala?Gly?Tyr?Asp?Ala?Leu?Arg?Leu?His?Thr?Glu
145?????????????????150?????????????????155?????????????????160
Ser?Arg?Met?Leu?Phe?Ala?Asp?Ala?Asp?Ser?Leu?Arg?Ile?Ser?Pro?Arg
165?????????????????170?????????????????175
Glu?Ala?Arg?Ser?Leu?Ile?Glu?Gln?Ala?Glu?Lys?Arg?Gln?Lys?Asp?Ala
180?????????????????185?????????????????190
Gln?Asn?Ala?Asp?Lys?Lys?Ala?Ala?Asp?Met?Leu?Ala?Glu?Tyr?Glu?Arg
195?????????????????200?????????????????205
Arg?Lys?Gly?Ile?Leu?Asp?Thr?Arg?Leu?Ser?Glu?Leu?Glu?Lys?Asn?Gly
210?????????????????215?????????????????220
Gly?Ala?Ala?Leu?Ala?Val?Leu?Asp?Ala?Gln?Gln?Ala?Arg?Leu?Leu?Gly
225?????????????????230?????????????????235?????????????????240
Gln?Gln?Thr?Arg?Asn?Asp?Arg?Ala?Ile?Ser?Glu?Ala?Arg?Asn?Lys?Leu
245?????????????????250?????????????????255
Ser?Ser?Val?Thr?Glu?Ser?Leu?Asn?Thr?Ala?Arg?Asn?Ala?Leu?Thr?Arg
260?????????????????265?????????????????270
Ala?Glu?Gln?Gln?Leu?Thr?Gln?Gln?Lys?Asn?Thr?Pro?Asp?Gly?Lys?Thr
275?????????????????280?????????????????285
Ile?Val?Ser?Pro?Glu?Lys?Phe?Pro?Gly?Arg?Ser?Ser?Thr?Asn?His?Ser
290?????????????????295?????????????????300
Ile?Val?Val?Ser?Gly?Asp?Pro?Arg?Phe?Ala?Gly?Thr?Ile?Lys?Ile?Thr
305?????????????????310?????????????????315?????????????????320
Thr?Ser?Ala?Val?Ile?Asp?Asn?Arg?Ala?Asn?Leu?Asn?Tyr?Leu?Leu?Ser
325?????????????????330?????????????????335
His?Ser?Gly?Leu?Asp?Tyr?Lys?Arg?Asn?Ile?Leu?Asn?Asp?Arg?Asn?Pro
340?????????????????345?????????????????350
Val?Val?Thr?Glu?Asp?Val?Glu?Gly?Asp?Lys?Lys?Ile?Tyr?Asn?Ala?Glu
355?????????????????360?????????????????365
Val?Ala?Glu?Trp?Asp?Lys?Leu?Arg?Gln?Arg?Leu?Leu?Asp?Ala?Arg?Asn
370?????????????????375?????????????????380
Lys?Ile?Thr?Ser?Ala?Glu?Ser?Ala?Val?Asn?Ser?Ala?Arg?Asn?Asn?Leu
385?????????????????390?????????????????395?????????????????400
Ser?Ala?Arg?Thr?Asn?Glu?Gln?Lys?His?Ala?Asn?Asp?Ala?Leu?Asn?Ala
405?????????????????410?????????????????415
Leu?Leu?Lys?Glu?Lys?Glu?Asn?Ile?Arg?Asn?Gln?Leu?Ser?Gly?Ile?Asn
420?????????????????425?????????????????430
Gln?Lys?Ile?Ala?Glu?Glu?Lys?Arg?Lys?Gln?Asp?Glu?Leu?Lys?Ala?Thr
435?????????????????440?????????????????445
Lys?Asp?Ala?Ile?Asn?Phe?Thr?Thr?Glu?Phe?Leu?Lys?Ser?Val?Ser?Glu
450?????????????????455?????????????????460
Lys?Tyr?Gly?Ala?Lys?Ala?Glu?Gln?Leu?Ala?Arg?Glu?Met?Ala?Gly?Gln
465?????????????????470?????????????????475?????????????????480
Ala?Lys?Gly?Lys?Lys?Ile?Arg?Asn?Val?Glu?Glu?Ala?Leu?Lys?Thr?Tyr
485?????????????????490?????????????????495
Glu?Lys?Tyr?Arg?Ala?Asp?Ile?Asn?Lys?Lys?Ile?Asn?Ala?Lys?Asp?Arg
500?????????????????505?????????????????510
Ala?Ala?Ile?Ala?Ala?Ala?Leu?Glu?Ser?Val?Lys?Leu?Ser?Asp?Ile?Ser
515?????????????????520?????????????????525
Ser?Asn?Leu?Asn?Arg?Phe?Ser?Arg?Gly?Leu?Gly?Tyr?Ala?Gly?Lys?Phe
530?????????????????535?????????????????540
Thr?Ser?Leu?Ala?Asp?Trp?Ile?Thr?Glu?Phe?Gly?Lys?Ala?Val?Arg?Thr
545?????????????????550?????????????????555?????????????????560
Glu?Asn?Trp?Arg?Pro?Leu?Phe?Val?Lys?Thr?Glu?Thr?Ile?Ile?Ala?Gly
565?????????????????570?????????????????575
Asn?Ala?Ala?Thr?Ala?Leu?Val?Ala?Leu?Val?Phe?Ser?Ile?Leu?Thr?Gly
580?????????????????585?????????????????590
Ser?Ala?Leu?Gly?Ile?Ile?Gly?Tyr?Gly?Leu?Leu?Met?Ala?Val?Thr?Gly
595?????????????????600?????????????????605
Ala?Leu?Ile?Asp?Glu?Ser?Leu?Val?Glu?Lys?Ala?Asn?Lys?Phe?Trp?Gly
610?????????????????615?????????????????620
Ile?Tyr?Ser?Thr?Cys?Tyr?Phe?Ile?Met
625?????????????????630
<210>20
<211>1905
<212>DNA
＜213〉artificial sequence
<220>
<221>CDS
<222>(4)..(1902)
＜223〉coding staphylococcus polypeptide
<400>20
atg?tct?gac?cct?gta?cgt?att?aca?aat?ccc?ggt?gca?gaa?tcg?ctg?ggg???????48
Ser?Asp?Pro?Val?Arg?Ile?Thr?Asn?Pro?Gly?Ala?Glu?Ser?Leu?Gly
1???????????????5???????????????????10??????????????????15
tat?gat?tca?gat?ggc?cat?gaa?att?atg?gcc?gtt?gat?att?tat?gta?aac???????96
Tyr?Asp?Ser?Asp?Gly?His?Glu?Ile?Met?Ala?Val?Asp?Ile?Tyr?Val?Asn
20??????????????????25??????????????????30
cct?cca?cgt?gtc?gat?gtc?ttt?cat?ggt?acc?ccg?cct?gca?tgg?agt?tcc??????144
Pro?Pro?Arg?Val?Asp?Val?Phe?His?Gly?Thr?Pro?Pro?Ala?Trp?Ser?Ser
35??????????????????40??????????????????45
ttc?ggg?aac?aaa?acc?atc?tgg?ggc?gga?aac?gag?tgg?gtt?gat?gat?tcc??????192
Phe?Gly?Asn?Lys?Thr?Ile?Trp?Gly?Gly?Asn?Glu?Trp?Val?Asp?Asp?Ser
50??????????????????55??????????????????60
cca?acc?cga?agt?gat?atc?gaa?aaa?agg?gac?aag?gaa?atc?aca?gcg?tac?????240
Pro?Thr?Arg?Ser?Asp?Ile?Glu?Lys?Arg?Asp?Lys?Glu?Ile?Thr?Ala?Tyr
65??????????????????70??????????????????75
aaa?aac?acg?ctc?agc?gcg?cag?cag?aaa?gag?aat?gag?aat?aag?cgt?act?????288
Lys?Asn?Thr?Leu?Ser?Ala?Gln?Gln?Lys?Glu?Asn?Glu?Asn?Lys?Arg?Thr
80??????????????????85??????????????????90??????????????????95
gaa?gcc?gga?aaa?cgc?ctc?tct?gcg?gcg?att?gct?gca?agg?gaa?aaa?gat?????336
Glu?Ala?Gly?Lys?Arg?Leu?Ser?Ala?Ala?Ile?Ala?Ala?Arg?Glu?Lys?Asp
100?????????????????105?????????????????110
gaa?aac?aca?ctg?aaa?aca?ctc?cgt?gcc?gga?aac?gca?gat?gcc?gct?gat?????384
Glu?Asn?Thr?Leu?Lys?Thr?Leu?Arg?Ala?Gly?Asn?Ala?Asp?Ala?Ala?Asp
115?????????????????120?????????????????125
att?aca?cga?cag?gag?ttc?aga?ctc?ctg?cag?gca?gag?ctg?aga?gaa?tac?????432
Ile?Thr?Arg?Gln?Glu?Phe?Arg?Leu?Leu?Gln?Ala?Glu?Leu?Arg?Glu?Tyr
130?????????????????135?????????????????140
gga?ttc?cgt?act?gaa?atc?gcc?gga?tat?gac?gcc?ctc?cgg?ctg?cat?aca?????480
Gly?Phe?Arg?Thr?Glu?Ile?Ala?Gly?Tyr?Asp?Ala?Leu?Arg?Leu?His?Thr
145?????????????????150?????????????????155
gag?agc?cgg?atg?ctg?ttt?gct?gat?gct?gat?tct?ctt?cgt?ata?tct?ccc?????528
Glu?Ser?Arg?Met?Leu?Phe?Ala?Asp?Ala?Asp?Ser?Leu?Arg?Ile?Ser?Pro
160?????????????????165?????????????????170?????????????????175
cgg?gag?gcc?agg?tcg?tta?atc?gaa?cag?gct?gaa?aaa?cgg?cag?aag?gat?????576
Arg?Glu?Ala?Arg?Ser?Leu?Ile?Glu?Gln?Ala?Glu?Lys?Arg?Gln?Lys?Asp
180?????????????????185?????????????????190
gcg?cag?aac?gca?gac?aag?aag?gcc?gct?gat?atg?ctt?gct?gaa?tac?gag?????624
Ala?Gln?Asn?Ala?Asp?Lys?Lys?Ala?Ala?Asp?Met?Leu?Ala?Glu?Tyr?Glu
195?????????????????200?????????????????205
cgc?aga?aaa?ggt?att?ctg?gac?acc?cgg?ttg?tca?gag?ctg?gaa?aaa?aat?????672
Arg?Arg?Lys?Gly?Ile?Leu?Asp?Thr?Arg?Leu?Ser?Glu?Leu?Glu?Lys?Asn
210?????????????????215?????????????????220
ggc?ggg?gca?gcc?ctt?gcc?gtt?ctt?gat?gca?caa?cag?gcc?cgt?ctg?ctc?????720
Gly?Gly?Ala?Ala?Leu?Ala?Val?Leu?Asp?Ala?Gln?Gln?Ala?Arg?Leu?Leu
225?????????????????230?????????????????235
ggg?cag?cag?aca?cgg?aat?gac?agg?gcc?att?tca?gag?gcc?cgg?aat?aaa?????768
Gly?Gln?Gln?Thr?Arg?Asn?Asp?Arg?Ala?Ile?Ser?Glu?Ala?Arg?Asn?Lys
240?????????????????245?????????????????250?????????????????255
ctc?agt?tca?gtg?acg?gaa?tcg?ctt?aac?acg?gcc?cgt?aat?gca?tta?acc?????816
Leu?Ser?Ser?Val?Thr?Glu?Ser?Leu?Asn?Thr?Ala?Arg?Asn?Ala?Leu?Thr
260?????????????????265?????????????????270
aga?gct?gaa?caa?cag?ctg?acg?caa?cag?aaa?aac?acg?cct?gac?ggc?aaa?????864
Arg?Ala?Glu?Gln?Gln?Leu?Thr?Gln?Gln?Lys?Asn?Thr?Pro?Asp?Gly?Lys
275?????????????????280?????????????????285
acg?ata?gtt?tcc?cct?gaa?aaa?ttc?ccg?ggg?cgt?tca?tca?aca?aat?cat?????912
Thr?Ile?Val?Ser?Pro?Glu?Lys?Phe?Pro?Gly?Arg?Ser?Ser?Thr?Asn?His
290?????????????????295?????????????????300
tct?att?gtt?gtg?agc?ggt?gat?ccg?aga?ttt?gcc?ggt?acg?ata?aaa?atc?????960
Ser?Ile?Val?Val?Ser?Gly?Asp?Pro?Arg?Phe?Ala?Gly?Thr?Ile?Lys?Ile
305?????????????????310?????????????????315
aca?acc?agc?gca?gtc?atc?gat?aac?cgt?gca?aac?ctg?aat?tat?ctt?ctg????1008
Thr?Thr?Ser?Ala?Val?Ile?Asp?Asn?Arg?Ala?Asn?Leu?Asn?Tyr?Leu?Leu
320?????????????????325?????????????????330?????????????????335
agc?cat?tcc?ggt?ctg?gac?tat?aaa?cgc?aat?att?ctg?aat?gac?cgg?aat????1056
Ser?His?Ser?Gly?Leu?Asp?Tyr?Lys?Arg?Asn?Ile?Leu?Asn?Asp?Arg?Asn
340?????????????????345?????????????????350
ccg?gtg?gtg?aca?gag?gat?gtg?gaa?ggt?gac?aag?aaa?att?tat?aat?gct????1104
Pro?Val?Val?Thr?Glu?Asp?Val?Glu?Gly?Asp?Lys?Lys?Ile?Tyr?Asn?Ala
355?????????????????360?????????????????365
gaa?gtt?gct?gaa?tgg?gat?aag?tta?cgg?caa?aga?ttg?ctt?gat?gcc?aga????1152
Glu?Val?Ala?Glu?Trp?Asp?Lys?Leu?Arg?Gln?Arg?Leu?Leu?Asp?Ala?Arg
370?????????????????375?????????????????380
aat?aaa?atc?acc?tct?gct?gaa?tct?gcg?gta?aat?tcg?gcg?aga?aat?aac?????1200
Asn?Lys?Ile?Thr?Ser?Ala?Glu?Ser?Ala?Val?Asn?Ser?Ala?Arg?Asn?Asn
385?????????????????390?????????????????395
ctc?agt?gcc?aga?aca?aat?gag?caa?aag?cat?gca?aat?gac?gct?ctt?aat?????1248
Leu?Ser?Ala?Arg?Thr?Asn?Glu?Gln?Lys?His?Ala?Asn?Asp?Ala?Leu?Asn
400?????????????????405?????????????????410?????????????????415
gcc?ctg?ttg?aag?gaa?aaa?gag?aat?ata?cgt?aac?cag?ctt?tcc?ggc?atc?????1296
Ala?Leu?Leu?Lys?Glu?Lys?Glu?Asn?Ile?Arg?Asn?Gln?Leu?Ser?Gly?Ile
420?????????????????425?????????????????430
aat?cag?aag?ata?gcg?gaa?gag?aaa?aga?aaa?cag?gat?gaa?ctg?aag?gca?????1344
Asn?Gln?Lys?Ile?Ala?Glu?Glu?Lys?Arg?Lys?Gln?Asp?Glu?Leu?Lys?Ala
435?????????????????440?????????????????445
acg?aaa?gac?gca?att?aat?ttc?aca?aca?gag?ttc?ctg?aaa?tca?gtt?tca?????1392
Thr?Lys?Asp?Ala?Ile?Asn?Phe?Thr?Thr?Glu?Phe?Leu?Lys?Ser?Val?Ser
450?????????????????455?????????????????460
gaa?aaa?tat?ggt?gca?aaa?gct?gag?cag?tta?gcc?aga?gag?atg?gcc?ggg?????1440
Glu?Lys?Tyr?Gly?Ala?Lys?Ala?Glu?Gln?Leu?Ala?Arg?Glu?Met?Ala?Gly
465?????????????????470?????????????????475
cag?gct?aaa?ggg?aag?aaa?ata?cgt?aat?gtt?gaa?gag?gca?tta?aaa?acg?????1488
Gln?Ala?Lys?Gly?Lys?Lys?Ile?Arg?Asn?Val?Glu?Glu?Ala?Leu?Lys?Thr
480?????????????????485?????????????????490?????????????????495
tat?gaa?aag?tac?cgg?gct?gac?att?aac?aaa?aaa?att?aat?gca?aaa?gat?????1536
Tyr?Glu?Lys?Tyr?Arg?Ala?Asp?Ile?Asn?Lys?Lys?Ile?Asn?Ala?Lys?Asp
500?????????????????505?????????????????510
cgt?gca?gcg?att?gcc?gca?gcc?ctt?gag?tct?gtg?aag?ctg?tct?gat?ata?????1584
Arg?Ala?Ala?Ile?Ala?Ala?Ala?Leu?Glu?Ser?Val?Lys?Leu?Ser?Asp?Ile
515?????????????????520?????????????????525
tcg?tct?aat?ctg?aac?aga?ttc?agt?cgg?gga?ctg?gga?tat?gca?gga?aaa????1632
Ser?Ser?Asn?Leu?Asn?Arg?Phe?Ser?Arg?Gly?Leu?Gly?Tyr?Ala?Gly?Lys
530?????????????????535?????????????????540
ttt?aca?agt?ctt?gct?gac?tgg?atc?act?gag?ttt?ggt?aag?gct?gtc?cgg?????1680
Phe?Thr?Ser?Leu?Ala?Asp?Trp?Ile?Thr?Glu?Phe?Gly?Lys?Ala?Val?Arg
545?????????????????550?????????????????555
aca?gag?aac?tgg?cgt?cct?ctt?ttt?gtt?aaa?aca?gaa?acc?atc?ata?gca?????1728
Thr?Glu?Asn?Trp?Arg?Pro?Leu?Phe?Val?Lys?Thr?Glu?Thr?Ile?Ile?Ala
560?????????????????565?????????????????570?????????????????575
ggc?aat?gcc?gca?acg?gct?ctt?gtg?gca?ctg?gtc?ttc?agt?att?ctt?acc?????1776
Gly?Asn?Ala?Ala?Thr?Ala?Leu?Val?Ala?Leu?Val?Phe?Ser?Ile?Leu?Thr
580?????????????????585?????????????????590
gga?agc?gct?tta?ggc?att?atc?ggg?tat?ggt?tta?ctg?atg?gct?gtc?acc?????1824
Gly?Ser?Ala?Leu?Gly?Ile?Ile?Gly?Tyr?Gly?Leu?Leu?Met?Ala?Val?Thr
595?????????????????600?????????????????605
ggt?gcg?ctg?att?gat?gaa?tcg?ctt?gtg?gaa?aaa?gcg?aat?aag?ttc?tgg?????1872
Gly?Ala?Leu?Ile?Asp?Glu?Ser?Leu?Val?Glu?Lys?Ala?Asn?Lys?Phe?Trp
610?????????????????615?????????????????620
ggt?att?gat?tcc?gtt?tgt?gca?tcc?tat?ttt?taa?????????????????????????1905
Gly?Ile?Asp?Ser?Val?Cys?Ala?Ser?Tyr?Phe
625?????????????????630
<210>21
<211>633
<212>PRT
＜213〉artificial sequence
<400>21
Ser?Asp?Pro?Val?Arg?Ile?Thr?Asn?Pro?Gly?Ala?Glu?Ser?Leu?Gly?Tyr
1???????????????5???????????????????10??????????????????15
Asp?Ser?Asp?Gly?His?Glu?Ile?Met?Ala?Val?Asp?Ile?Tyr?Val?Asn?Pro
20??????????????????25??????????????????30
Pro?Arg?Val?Asp?Val?Phe?His?Gly?Thr?Pro?Pro?Ala?Trp?Ser?Ser?Phe
35??????????????????40??????????????????45
Gly?Asn?Lys?Thr?Ile?Trp?Gly?Gly?Asn?Glu?Trp?Val?Asp?Asp?Ser?Pro
50?????????????????55??????????????????60
Thr?Arg?Ser?Asp?Ile?Glu?Lys?Arg?Asp?Lys?Glu?Ile?Thr?Ala?Tyr?Lys
65??????????????????70??????????????????75??????????????????80
Asn?Thr?Leu?Ser?Ala?Gln?Gln?Lys?Glu?Asn?Glu?Asn?Lys?Arg?Thr?Glu
85??????????????????90??????????????????95
Ala?Gly?Lys?Arg?Leu?Ser?Ala?Ala?Ile?Ala?Ala?Arg?Glu?Lys?Asp?Glu
100?????????????????105?????????????????110
Asn?Thr?Leu?Lys?Thr?Leu?Arg?Ala?Gly?Asn?Ala?Asp?Ala?Ala?Asp?Ile
115?????????????????120?????????????????125
Thr?Arg?Gln?Glu?Phe?Arg?Leu?Leu?Gln?Ala?Glu?Leu?Arg?Glu?Tyr?Gly
130?????????????????135?????????????????140
Phe?Arg?Thr?Glu?Ile?Ala?Gly?Tyr?Asp?Ala?Leu?Arg?Leu?His?Thr?Glu
145?????????????????150?????????????????155?????????????????160
Ser?Arg?Met?Leu?Phe?Ala?Asp?Ala?Asp?Ser?Leu?Arg?Ile?Ser?Pro?Arg
165?????????????????170?????????????????175
Glu?Ala?Arg?Ser?Leu?Ile?Glu?Gln?Ala?Glu?Lys?Arg?Gln?Lys?Asp?Ala
180?????????????????185?????????????????190
Gln?Asn?Ala?Asp?Lys?Lys?Ala?Ala?Asp?Met?Leu?Ala?Glu?Tyr?Glu?Arg
195?????????????????200?????????????????205
Arg?Lys?Gly?Ile?Leu?Asp?Thr?Arg?Leu?Ser?Glu?Leu?Glu?Lys?Asn?Gly
210?????????????????215?????????????????220
Gly?Ala?Ala?Leu?Ala?Val?Leu?Asp?Ala?Gln?Gln?Ala?Arg?Leu?Leu?Gly
225?????????????????230?????????????????235?????????????????240
Gln?Gln?Thr?Arg?Asn?Asp?Arg?Ala?Ile?Ser?Glu?Ala?Arg?Asn?Lys?Leu
245?????????????????250?????????????????255
Ser?Ser?Val?Thr?Glu?Ser?Leu?Asn?Thr?Ala?Arg?Asn?Ala?Leu?Thr?Arg
260?????????????????265?????????????????270
Ala?Glu?Gln?Gln?Leu?Thr?Gln?Gln?Lys?Asn?Thr?Pro?Asp?Gly?Lys?Thr
275?????????????????280?????????????????285
Ile?Val?Ser?Pro?Glu?Lys?Phe?Pro?Gly?Arg?Ser?Ser?Thr?Asn?His?Ser
290?????????????????295?????????????????300
Ile?Val?Val?Ser?Gly?Asp?Pro?Arg?Phe?Ala?Gly?Thr?Ile?Lys?Ile?Thr
305?????????????????310?????????????????315?????????????????320
Thr?Ser?Ala?Val?Ile?Asp?Asn?Arg?Ala?Asn?Leu?Asn?Tyr?Leu?Leu?Ser
325?????????????????330?????????????????335
His?Ser?Gly?Leu?Asp?Tyr?Lys?Arg?Asn?Ile?Leu?Asn?Asp?Arg?Asn?Pro
340?????????????????345?????????????????350
Val?Val?Thr?Glu?Asp?Val?Glu?Gly?Asp?Lys?Lys?Ile?Tyr?Asn?Ala?Glu
355?????????????????360?????????????????365
Val?Ala?Glu?Trp?Asp?Lys?Leu?Arg?Gln?Arg?Leu?Leu?Asp?Ala?Arg?Asn
370?????????????????375?????????????????380
Lys?Ile?Thr?Ser?Ala?Glu?Ser?Ala?Val?Asn?Ser?Ala?Arg?Asn?Asn?Leu
385?????????????????390?????????????????395?????????????????400
Ser?Ala?Arg?Thr?Asn?Glu?Gln?Lys?His?Ala?Asn?Asp?Ala?Leu?Asn?Ala
405?????????????????410?????????????????415
Leu?Leu?Lys?Glu?Lys?Glu?Asn?Ile?Arg?Asn?Gln?Leu?Ser?Gly?Ile?Asn
420?????????????????425?????????????????430
Gln?Lys?Ile?Ala?Glu?Glu?Lys?Arg?Lys?Gln?Asp?Glu?Leu?Lys?Ala?Thr
435?????????????????440?????????????????445
Lys?Asp?Ala?Ile?Asn?Phe?Thr?Thr?Glu?Phe?Leu?Lys?Ser?Val?Ser?Glu
450?????????????????455?????????????????460
Lys?Tyr?Gly?Ala?Lys?Ala?Glu?Gln?Leu?Ala?Arg?Glu?Met?Ala?Gly?Gln
465?????????????????470?????????????????475?????????????????480
Ala?Lys?Gly?Lys?Lys?Ile?Arg?Asn?Val?Glu?Glu?Ala?Leu?Lys?Thr?Tyr
485?????????????????490?????????????????495
Glu?Lys?Tyr?Arg?Ala?Asp?Ile?Asn?Lys?Lys?Ile?Asn?Ala?Lys?Asp?Arg
500?????????????????505?????????????????510
Ala?Ala?Ile?Ala?Ala?Ala?Leu?Glu?Ser?Val?Lys?Leu?Ser?Asp?Ile?Ser
515?????????????????520?????????????????525
Ser?Asn?Leu?Asn?Arg?Phe?Ser?Arg?Gly?Leu?Gly?Tyr?Ala?Gly?Lys?Phe
530?????????????????535?????????????????540
Thr?Ser?Leu?Ala?Asp?Trp?Ile?Thr?Glu?Phe?Gly?Lys?Ala?Val?Arg?Thr
545?????????????????550?????????????????555?????????????????560
Glu?Asn?Trp?Arg?Pro?Leu?Phe?Val?Lys?Thr?Glu?Thr?Ile?Ile?Ala?Gly
565?????????????????570?????????????????575
Asn?Ala?Ala?Thr?Ala?Leu?Val?Ala?Leu?Val?Phe?Ser?Ile?Leu?Thr?Gly
580?????????????????585?????????????????590
Ser?Ala?Leu?Gly?Ile?Ile?Gly?Tyr?Gly?Leu?Leu?Met?Ala?Val?Thr?Gly
595?????????????????600?????????????????605
Ala?Leu?Ile?Asp?Glu?Ser?Leu?Val?Glu?Lys?Ala?Asn?Lys?Phe?Trp?Gly
610?????????????????615?????????????????620
Ile?Asp?Ser?Val?Cys?Ala?Ser?Tyr?Phe
625?????????????????630
<210>22
<211>1905
<212>DNA
＜213〉artificial sequence
<220>
<221>CDS
<222>(4)..(1902)
＜223〉coding staphylococcus polypeptide
<400>22
atg?tat?tcc?acc?tgt?gat?ttt?ata?atg?tct?gac?cct?gta?cgt?att?aca?????48
Tyr?Ser?Thr?Cys?Asp?Phe?Ile?Met?Ser?Asp?Pro?Val?Arg?Ile?Thr
1???????????????5???????????????????10??????????????????15
aat?ccc?ggt?gca?gaa?tcg?ctg?ggg?tat?gat?tca?gat?ggc?cat?gaa?att?????96
Asn?Pro?Gly?Ala?Glu?Ser?Leu?Gly?Tyr?Asp?Ser?Asp?Gly?His?Glu?Ile
20??????????????????25??????????????????30
atg?gcc?gtt?gat?att?tat?gta?aac?cct?cca?cgt?gtc?gat?gtc?ttt?cat????144
Met?Ala?Val?Asp?Ile?Tyr?Val?Asn?Pro?Pro?Arg?Val?Asp?Val?Phe?His
35??????????????????40??????????????????45
ggt?acc?ccg?cct?gca?tgg?agt?tcc?ttc?ggg?aac?aaa?acc?atc?tgg?ggc????192
Gly?Thr?Pro?Pro?Ala?Trp?Ser?Ser?Phe?Gly?Asn?Lys?Thr?Ile?Trp?Gly
50??????????????????55??????????????????60
gga?aac?gag?tgg?gtt?gat?gat?tcc?cca?acc?cga?agt?gat?atc?gaa?aaa????240
Gly?Asn?Glu?Trp?Val?Asp?Asp?Ser?Pro?Thr?Arg?Ser?Asp?Ile?Glu?Lys
65??????????????????70??????????????????75
agg?gac?aag?gaa?atc?aca?gcg?tac?aaa?aac?acg?ctc?agc?gcg?cag?cag????288
Arg?Asp?Lys?Glu?Ile?Thr?Ala?Tyr?Lys?Asn?Thr?Leu?Ser?Ala?Gln?Gln
80??????????????????85??????????????????90??????????????????95
aaa?gag?aat?gag?aat?aag?cgt?act?gaa?gcc?gga?aaa?cgc?ctc?tct?gcg????336
Lys?Glu?Asn?Glu?Asn?Lys?Arg?Thr?Glu?Ala?Gly?Lys?Arg?Leu?Ser?Ala
100?????????????????105?????????????????110
gcg?att?gct?gca?agg?gaa?aaa?gat?gaa?aac?aca?ctg?aaa?aca?ctc?cgt????384
Ala?Ile?Ala?Ala?Arg?Glu?Lys?Asp?Glu?Asn?Thr?Leu?Lys?Thr?Leu?Arg
115?????????????????120?????????????????125
gcc?gga?aac?gca?gat?gcc?gct?gat?att?aca?cga?cag?gag?ttc?aga?ctc????432
Ala?Gly?Asn?Ala?Asp?Ala?Ala?Asp?Ile?Thr?Arg?Gln?Glu?Phe?Arg?Leu
130?????????????????135?????????????????140
ctg?cag?gca?gag?ctg?aga?gaa?tac?gga?ttc?cgt?act?gaa?atc?gcc?gga????480
Leu?Gln?Ala?Glu?Leu?Arg?Glu?Tyr?Gly?Phe?Arg?Thr?Glu?Ile?Ala?Gly
145?????????????????150?????????????????155
tat?gac?gcc?ctc?cgg?ctg?cat?aca?gag?agc?cgg?atg?ctg?ttt?gct?gat????528
Tyr?Asp?Ala?Leu?Arg?Leu?His?Thr?Glu?Ser?Arg?Met?Leu?Phe?Ala?Asp
160?????????????????165?????????????????170?????????????????175
gct?gat?tct?ctt?cgt?ata?tct?ccc?cgg?gag?gcc?agg?tcg?tta?atc?gaa????576
Ala?Asp?Ser?Leu?Arg?Ile?Ser?Pro?Arg?Glu?Ala?Arg?Ser?Leu?Ile?Glu
180?????????????????185?????????????????190
cag?gct?gaa?aaa?cgg?cag?aag?gat?gcg?cag?aac?gca?gac?aag?aag?gcc????624
Gln?Ala?Glu?Lys?Arg?Gln?Lys?Asp?Ala?Gln?Asn?Ala?Asp?Lys?Lys?Ala
195?????????????????200?????????????????205
gct?gat?atg?ctt?gct?gaa?tac?gag?cgc?aga?aaa?ggt?att?ctg?gac?acc????672
Ala?Asp?Met?Leu?Ala?Glu?Tyr?Glu?Arg?Arg?Lys?Gly?Ile?Leu?Asp?Thr
210?????????????????215?????????????????220
cgg?ttg?tca?gag?ctg?gaa?aaa?aat?ggc?ggg?gca?gcc?ctt?gcc?gtt?ctt????720
Arg?Leu?Ser?Glu?Leu?Glu?Lys?Asn?Gly?Gly?Ala?Ala?Leu?Ala?Val?Leu
225?????????????????230?????????????????235
gat?gca?caa?cag?gcc?cgt?ctg?ctc?ggg?cag?cag?aca?cgg?aat?gac?agg????768
Asp?Ala?Gln?Gln?Ala?arg?Leu?Leu?Gly?Gln?Gln?Thr?Arg?Asn?Asp?Arg
240?????????????????245?????????????????250?????????????????255
gcc?att?tca?gag?gcc?cgg?aat?aaa?ctc?agt?tca?gtg?acg?gaa?tcg?ctt??????816
Ala?Ile?Ser?Glu?Ala?arg?Asn?Lys?Leu?Ser?Ser?Val?Thr?Glu?Ser?Leu
260?????????????????265?????????????????270
aac?acg?gcc?cgt?aat?gca?tta?acc?aga?gct?gaa?caa?cag?ctg?acg?caa??????864
Asn?Thr?Ala?Arg?Asn?ala?Leu?Thr?Arg?Ala?Glu?Gln?Gln?Leu?Thr?Gln
275?????????????????280?????????????????285
cag?aaa?aac?acg?cct?gac?ggc?aaa?acg?ata?gtt?tcc?cct?gaa?aaa?ttc??????912
Gln?Lys?Asn?Thr?Pro?Asp?Gly?Lys?Thr?Ile?Val?Ser?Pro?Glu?Lys?Phe
290?????????????????295?????????????????300
ccg?ggg?cgt?tca?tca?aca?aat?cat?tct?att?gtt?gtg?agc?ggt?gat?ccg??????960
Pro?Gly?Arg?Ser?Ser?Thr?Asn?His?Ser?Ile?Val?Val?Ser?Gly?Asp?Pro
305?????????????????310?????????????????315
aga?ttt?gcc?ggt?acg?ata?aaa?atc?aca?acc?agc?gca?gtc?atc?gat?aac?????1008
Arg?Phe?Ala?Gly?Thr?Ile?Lys?Ile?Thr?Thr?Ser?Ala?Val?Ile?Asp?Asn
320?????????????????325?????????????????330?????????????????335
cgt?gca?aac?ctg?aat?tat?ctt?ctg?agc?cat?tcc?ggt?ctg?gac?tat?aaa?????1056
Arg?Ala?Asn?Leu?Asn?Tyr?Leu?Leu?Ser?His?Ser?Gly?Leu?Asp?Tyr?Lys
340?????????????????345?????????????????350
cgc?aat?att?ctg?aat?gac?cgg?aat?ccg?gtg?gtg?aca?gag?gat?gtg?gaa?????1104
Arg?Asn?Ile?Leu?Asn?Asp?Arg?Asn?Pro?Val?Val?Thr?Glu?Asp?Val?Glu
355?????????????????360?????????????????365
ggt?gac?aag?aaa?att?tat?aat?gct?gaa?gtt?gct?gaa?tgg?gat?aag?tta?????1152
Gly?Asp?Lys?Lys?Ile?Tyr?Asn?Ala?Glu?Val?Ala?Glu?Trp?Asp?Lys?Leu
370?????????????????375?????????????????380
cgg?caa?aga?ttg?ctt?gat?gcc?aga?aat?aaa?atc?acc?tct?gct?gaa?tct?????1200
Arg?Gln?Arg?Leu?Leu?Asp?Ala?Arg?Asn?Lys?Ile?Thr?Ser?Ala?Glu?Ser
385?????????????????390?????????????????395
gcg?gta?aat?tcg?gcg?aga?aat?aac?ctc?agt?gcc?aga?aca?aat?gag?caa?????1248
Ala?Val?Asn?Ser?Ala?Arg?Asn?Asn?Leu?Ser?Ala?Arg?Thr?Asn?Glu?Gln
400?????????????????405?????????????????410?????????????415
aag?cat?gca?aat?gac?gct?ctt?aat?gcc?ctg?ttg?aag?gaa?aaa?gag?aat?????1296
Lys?His?Ala?Asn?Asp?Ala?Leu?Asn?Ala?Leu?Leu?Lys?Glu?Lys?Glu?Asn
420?????????????????425?????????????????430
ata?cgt?aac?cag?ctt?tcc?ggc?atc?aat?cag?aag?ata?gcg?gaa?gag?aaa?????1344
Ile?Arg?Asn?Gln?Leu?Ser?Gly?Ile?Asn?Gln?Lys?Ile?Ala?Glu?Glu?Lys
435?????????????????440?????????????????445
aga?aaa?cag?gat?gaa?ctg?aag?gca?acg?aaa?gac?gca?att?aat?ttc?aca?????1392
Arg?Lys?Gln?Asp?Glu?Leu?Lys?Ala?Thr?Lys?Asp?Ala?Ile?Asn?Phe?Thr
450?????????????????455?????????????????460
aca?gag?ttc?ctg?aaa?tca?gtt?tca?gaa?aaa?tat?ggt?gca?aaa?gct?gag?????1440
Thr?Glu?Phe?Leu?Lys?Ser?Val?Ser?Glu?Lys?Tyr?Gly?Ala?Lys?Ala?Glu
465?????????????????470?????????????????475
cag?tta?gcc?aga?gag?atg?gcc?ggg?cag?gct?aaa?ggg?aag?aaa?ata?cgt?????1488
Gln?Leu?Ala?Arg?Glu?Met?Ala?Gly?Gln?Ala?Lys?Gly?Lys?Lys?Ile?Arg
480?????????????????485?????????????????490?????????????????495
aat?gtt?gaa?gag?gca?tta?aaa?acg?tat?gaa?aag?tac?cgg?gct?gac?att?????1536
Asn?Val?Glu?Glu?Ala?Leu?Lys?Thr?Tyr?Glu?Lys?Tyr?Arg?Ala?Asp?Ile
500?????????????????505?????????????????510
aac?aaa?aaa?att?aat?gca?aaa?gat?cgt?gca?gcg?att?gcc?gca?gcc?ctt?????1584
Asn?Lys?Lys?Ile?Asn?Ala?Lys?Asp?Arg?Ala?Ala?Ile?Ala?Ala?Ala?Leu
515?????????????????520????????????????525
gag?tct?gtg?aag?ctg?tct?gat?ata?tcg?tct?aat?ctg?aac?aga?ttc?agt?????1632
Glu?Ser?Val?Lys?Leu?Ser?Asp?Ile?Ser?Ser?Asn?Leu?Asn?Arg?Phe?Ser
530?????????????????535?????????????????540
cgg?gga?ctg?gga?tat?gca?gga?aaa?ttt?aca?agt?ctt?gct?gac?tgg?atc?????1680
Arg?Gly?Leu?Gly?Tyr?Ala?Gly?Lys?Phe?Thr?Ser?Leu?Ala?Asp?Trp?Ile
545?????????????????550?????????????????555
act?gag?ttt?ggt?aag?gct?gtc?cgg?aca?gag?aac?tgg?cgt?cct?ctt?ttt?????1728
Thr?Glu?Phe?Gly?Lys?Ala?Val?Arg?Thr?Glu?Asn?Trp?Arg?Pro?Leu?Phe
560?????????????????565?????????????????570?????????????????575
gtt?aaa?aca?gaa?acc?atc?ata?gca?ggc?aat?gcc?gca?acg?gct?ctt?gtg?????1776
Val?Lys?Thr?Glu?Thr?Ile?Ile?Ala?Gly?Asn?Ala?Ala?Thr?Ala?Leu?Val
580?????????????????585?????????????????590
gca?ctg?gtc?ttc?agt?att?ctt?acc?gga?agc?gct?tta?ggc?att?atc?ggg?????1824
Ala?Leu?Val?Phe?Ser?Ile?Leu?Thr?Gly?Ser?Ala?Leu?Gly?Ile?Ile?Gly
595?????????????????600?????????????????605
tat?ggt?tta?ctg?atg?gct?gtc?acc?ggt?gcg?ctg?att?gat?gaa?tcg?ctt?????1872
Tyr?Gly?Leu?Leu?Met?Ala?Val?Thr?Gly?Ala?Leu?Ile?Asp?Glu?Ser?Leu
610?????????????????615?????????????????620
gtg?gaa?aaa?gcg?aat?aag?ttc?tgg?ggt?att?taa?????????????????????????1905
Val?Glu?Lys?Ala?Asn?Lys?Phe?Trp?Gly?Ile
625?????????????????630
<210>23
<211>633
<212>PRT
＜213〉artificial sequence
<400>23
Tyr?Ser?Thr?Cys?Asp?Phe?Ile?Met?Ser?Asp?Pro?Val?Arg?Ile?Thr?Asn
1???????????????5???????????????????10??????????????????15
Pro?Gly?Ala?Glu?Ser?Leu?Gly?Tyr?Asp?Ser?Asp?Gly?His?Glu?Ile?Met
20??????????????????25??????????????????30
Ala?Val?Asp?Ile?Tyr?Val?Asn?Pro?Pro?Arg?Val?Asp?Val?Phe?His?Gly
35??????????????????40??????????????????45
Thr?Pro?Pro?Ala?Trp?Ser?Ser?Phe?Gly?Asn?Lys?Thr?Ile?Trp?Gly?Gly
50??????????????????55??????????????????60
Asn?Glu?Trp?Val?Asp?Asp?Ser?Pro?Thr?Arg?Ser?Asp?Ile?Glu?Lys?Arg
65??????????????????70??????????????????75??????????????????80
Asp?Lys?Glu?Ile?Thr?Ala?Tyr?Lys?Asn?Thr?Leu?Ser?Ala?Gln?Gln?Lys
85??????????????????90??????????????????95
Glu?Asn?Glu?Asn?Lys?Arg?Thr?Glu?Ala?Gly?Lys?Arg?Leu?Ser?Ala?Ala
100?????????????????105?????????????????110
Ile?Ala?Ala?Arg?Glu?Lys?Asp?Glu?Asn?Thr?Leu?Lys?Thr?Leu?Arg?Ala
115?????????????????120?????????????????125
Gly?Asn?Ala?Asp?Ala?Ala?Asp?Ile?Thr?Arg?Gln?Glu?Phe?Arg?Leu?Leu
130?????????????????135?????????????????140
Gln?Ala?Glu?Leu?Arg?Glu?Tyr?Gly?Phe?Arg?Thr?Glu?Ile?Ala?Gly?Tyr
145?????????????????150?????????????????155?????????????????160
Asp?Ala?Leu?Arg?Leu?His?Thr?Glu?Ser?Arg?Met?Leu?Phe?Ala?Asp?Ala
165?????????????????170?????????????????175
Asp?Ser?Leu?Arg?Ile?Ser?Pro?Arg?Glu?Ala?Arg?Ser?Leu?Ile?Glu?Gln
180?????????????????185?????????????????190
Ala?Glu?Lys?Arg?Gln?Lys?Asp?Ala?Gln?Asn?Ala?Asp?Lys?Lys?Ala?Ala
195?????????????????200?????????????????205
Asp?Met?Leu?Ala?Glu?Tyr?Glu?Arg?Arg?Lys?Gly?Ile?Leu?Asp?Thr?Arg
210?????????????????215?????????????????220
Leu?Ser?Glu?Leu?Glu?Lys?Asn?Gly?Gly?Ala?Ala?Leu?Ala?Val?Leu?Asp
225?????????????????230?????????????????235?????????????????240
Ala?Gln?Gln?Ala?Arg?Leu?Leu?Gly?Gln?Gln?Thr?Arg?Asn?Asp?Arg?Ala
245?????????????????250?????????????????255
Ile?Ser?Glu?Ala?Arg?Asn?Lys?Leu?Ser?Ser?Val?Thr?Glu?Ser?Leu?Asn
260?????????????????265?????????????????270
Thr?Ala?Arg?Asn?Ala?Leu?Thr?Arg?Ala?Glu?Gln?Gln?Leu?Thr?Gln?Gln
275?????????????????280?????????????????285
Lys?Asn?Thr?Pro?Asp?Gly?Lys?Thr?Ile?Val?Ser?Pro?Glu?Lys?Phe?Pro
290?????????????????295?????????????????300
Gly?Arg?Ser?Ser?Thr?Asn?His?Ser?Ile?Val?Val?Ser?Gly?Asp?Pro?Arg
305?????????????????310?????????????????315?????????????????320
Phe?Ala?Gly?Thr?Ile?Lys?Ile?Thr?Thr?Ser?Ala?Val?Ile?Asp?Asn?Arg
325?????????????????330?????????????????335
Ala?Asn?Leu?Asn?Tyr?Leu?Leu?Ser?His?Ser?Gly?Leu?Asp?Tyr?Lys?Arg
340?????????????????345?????????????????350
Asn?Ile?Leu?Asn?Asp?Arg?Asn?Pro?Val?Val?Thr?Glu?Asp?Val?Glu?Gly
355?????????????????360?????????????????365
Asp?Lys?Lys?Ile?Tyr?Asn?Ala?Glu?Val?Ala?Glu?Trp?Asp?Lys?Leu?Arg
370?????????????????375?????????????????380
Gln?Arg?Leu?Leu?Asp?Ala?Arg?Asn?Lys?Ile?Thr?Ser?Ala?Glu?Ser?Ala
385?????????????????390?????????????????395?????????????????400
Val?Asn?Ser?Ala?Arg?Asn?Asn?Leu?Ser?Ala?Arg?Thr?Asn?Glu?Gln?Lys
405?????????????????410?????????????????415
His?Ala?Asn?Asp?Ala?Leu?Asn?Ala?Leu?Leu?Lys?Glu?Lys?Glu?Asn?Ile
420?????????????????425?????????????????430
Arg?Asn?Gln?Leu?Ser?Gly?Ile?Asn?Gln?Lys?Ile?Ala?Glu?Glu?Lys?Arg
435?????????????????440?????????????????445
Lys?Gln?Asp?Glu?Leu?Lys?Ala?Thr?Lys?Asp?Ala?Ile?Asn?Phe?Thr?Thr
450?????????????????455?????????????????460
Glu?Phe?Leu?Lys?Ser?Val?Ser?Glu?Lys?Tyr?Gly?Ala?Lys?Ala?Glu?Gln
465?????????????????470?????????????????475?????????????????480
Leu?Ala?Arg?Glu?Met?Ala?Gly?Gln?Ala?Lys?Gly?Lys?Lys?Ile?Arg?Asn
485?????????????????490?????????????????495
Val?Glu?Glu?Ala?Leu?Lys?Thr?Tyr?Glu?Lys?Tyr?Arg?Ala?Asp?Ile?Asn
500?????????????????505?????????????????510
Lys?Lys?Ile?Asn?Ala?Lys?Asp?Arg?Ala?Ala?Ile?Ala?Ala?Ala?Leu?Glu
515?????????????????520?????????????????525
Ser?Val?Lys?Leu?Ser?Asp?Ile?Ser?Ser?Asn?Leu?Asn?Arg?Phe?Ser?Arg
530?????????????????535?????????????????540
Gly?Leu?Gly?Tyr?Ala?Gly?Lys?Phe?Thr?Ser?Leu?Ala?Asp?Trp?Ile?Thr
545?????????????????550?????????????????555?????????????????560
Glu?Phe?Gly?Lys?Ala?Val?Arg?Thr?Glu?Asn?Trp?Arg?Pro?Leu?Phe?Val
565?????????????????570?????????????????575
Lys?Thr?Glu?Thr?Ile?Ile?Ala?Gly?Asn?Ala?Ala?Thr?Ala?Leu?Val?Ala
580?????????????????585?????????????????590
Leu?Val?Phe?Ser?Ile?Leu?Thr?Gly?Ser?Ala?Leu?Gly?Ile?Ile?Gly?Tyr
595?????????????????600?????????????????605
Gly?Leu?Leu?Met?Ala?Val?Thr?Gly?Ala?Leu?Ile?Asp?Glu?Ser?Leu?Val
610?????????????????615?????????????????620
Glu?Lys?Ala?Asn?Lys?Phe?Trp?Gly?Ile
625?????????????????630
<210>24
<211>66
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
<222>(1)..(66)
＜223〉primer
<400>24
gcgaataagt?tctggggtat?ttattccacc?tgtgatttta?taatgtaaat?aaaatataag????60
acaggc???????????????????????????????????????????????????????????????66
<210>25
<211>66
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
<222>(1)..(66)
＜223〉primer
<400>25
gcctgtctta?tattttattt?acattataaa?atcacaggtg?gaataaatac?cccagaactt????60
attcgc???????????????????????????????????????????????????????????????66
<210>26
<211>69
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
<222>(1)..(69)
＜223〉primer
<400>26
gcgaataagt?tctggggtat?tggagttaac?gcatgttctt?ccctgtttta?aataaaatat????60
aagacaggc????????????????????????????????????????????????????????????69
<210>27
<211>69
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
<222>(1)..(69)
＜223〉primer
<400>27
gcctgtctta?tattttattt?aaaacaggga?agaacatgcg?ttaactccaa?taccccagaa????60
cttattcgc????????????????????????????????????????????????????????????69
<210>28
<211>66
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
<222>(1)..(66)
＜223〉primer
<400>28
gcgaataagt?tctggggtat?ttatataaac?tgtgattttc?ttctgtaaat?aaaatataag????60
acaggc???????????????????????????????????????????????????????????????66
<210>29
<211>66
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
<222>(1)..(66)
＜223〉primer
<400>29
gcctgtctta?tattttattt?acagaagaaa?atcacagttt?atataaatac?cccagaactt????60
attcgc???????????????????????????????????????????????????????????????66
<210>30
<211>66
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
<222>(1)..(66)
＜223〉primer
<400>30
gcgaataagt?tctggggtat?ttattccacc?tgttacttta?taatgtaaat?aaaatataag????60
acaggc???????????????????????????????????????????????????????????????66
<210>31
<211>66
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
<222>(1)..(66)
＜223〉primer
<400>31
gcctgtctta?tattttattt?acattataaa?gtaacaggtg?gaataaatac?cccagaactt????60
attcgc???????????????????????????????????????????????????????????????66
<210>32
<211>66
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
<222>(1)..(66)
＜223〉primer
<400>32
gcgaataagt?tctggggtat?tgattccgtt?tgtgcatcct?atttttaaat?aaaatataag????60
acaggc???????????????????????????????????????????????????????????????66
<210>33
<211>66
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
<222>(1)..(66)
＜223〉primer
<400>33
gcctgtctta?tattttattt?aaaaatagga?tgcacaaacg?gaatcaatac?cccagaactt????60
attcgc???????????????????????????????????????????????????????????????66

Claims (27)

1. the Nucleotide of coding reorganization antimicrobial polypeptide is characterized in that it contains
Coding can form the colicin of ionic channel or the gene in its water-based pore passage structure territory, and
The gene of coding staphylococcus signal conduction polypeptide.
2. Nucleotide according to claim 1, the gene of wherein said coding staphylococcus signal conduction polypeptide are connected in the C-terminal that described coding can form the gene in the colicin of ionic channel or its water-based pore passage structure territory.
3. Nucleotide according to claim 1, the gene of wherein said coding staphylococcus signal conduction polypeptide are connected in the N-terminal that described coding can form the gene in the colicin of ionic channel or its water-based pore passage structure territory.
4. according to claim 2 or 3 described Nucleotide, the wherein said colicin that forms ionic channel is selected from colicin E1, Ia, Ib, A, B and N.
5. Nucleotide according to claim 4, the wherein said colicin that forms ionic channel is colicin Ia.
6. Nucleotide according to claim 5 wherein contains the nucleotide sequence of SEQ ID NO.12.
7. Nucleotide according to claim 5 wherein contains the nucleotide sequence of SEQ ID NO.14.
8. Nucleotide according to claim 5 wherein contains the nucleotide sequence of SEQ ID NO.16.
9. Nucleotide according to claim 5 wherein contains the nucleotide sequence of SEQ ID NO.18.
10. Nucleotide according to claim 5 wherein contains the nucleotide sequence of SEQ ID NO.20.
11. Nucleotide according to claim 5 wherein contains the nucleotide sequence of SEQ ID NO.22.
12. a recombinant plasmid is characterized in that it contains
Coding can form the colicin of ionic channel or the gene in its water-based pore passage structure territory, and
The gene of coding staphylococcus signal conduction polypeptide.
13. recombinant plasmid according to claim 12 wherein contains the nucleotide sequence of SEQ ID NO.12.
14. recombinant plasmid according to claim 12 wherein contains the nucleotide sequence of SEQ ID NO.14.
15. recombinant plasmid according to claim 12 wherein contains the nucleotide sequence of SEQ ID NO.16.
16. recombinant plasmid according to claim 12 wherein contains the nucleotide sequence of SEQ ID NO.18.
17. recombinant plasmid according to claim 12 wherein contains the nucleotide sequence of SEQ ID NO.20.
18. recombinant plasmid according to claim 12 wherein contains the nucleotide sequence of SEQ ID NO.22.
19. a reorganization antimicrobial polypeptide, it contains
Can form colicin or its water-based pore passage structure territory of ionic channel, and
Staphylococcus signal conduction polypeptide.
20. antimicrobial polypeptide according to claim 19 wherein comprises the aminoacid sequence of SEQ ID NO.13.
21. antimicrobial polypeptide according to claim 19 wherein comprises the aminoacid sequence of SEQ ID NO.15.
22. antimicrobial polypeptide according to claim 19 wherein comprises the aminoacid sequence of SEQ ID NO.17.
23. antimicrobial polypeptide according to claim 19 wherein comprises the aminoacid sequence of SEQ ID NO.19.
24. antimicrobial polypeptide according to claim 19 wherein comprises the aminoacid sequence of SEQ ID NO.21.
25. antimicrobial polypeptide according to claim 19 wherein comprises the aminoacid sequence of SEQ ID NO.23.
26. contain the pharmaceutical composition of any one described reorganization antimicrobial polypeptide of claim 20～25.
27. the preparation method of reorganization antimicrobial polypeptide may further comprise the steps:
The gene that the gene of coding staphylococcus signal conduction polypeptide and coding can be formed the colicin of ionic channel or its water-based pore passage structure territory is operably connected and obtains the recombinate gene of antimicrobial polypeptide of coding;
The gene that obtains imported in the expression system express;
Separate polypeptide expressed and obtain antimicrobial polypeptide of the present invention.
CNA031204007A 2002-11-12 2003-03-19 Novel antibiotic polypeptide and its preparing method Withdrawn CN1532284A (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
CNA031204007A CN1532284A (en) 2002-11-12 2003-03-19 Novel antibiotic polypeptide and its preparing method
PCT/CN2004/000046 WO2004082701A1 (en) 2003-03-19 2004-01-14 New engineered antibiotic peptide and its preparing process

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CNA021340420A CN1513877A (en) 2002-11-12 2002-11-12 Artificial combined antistaphylococcus engineering multipeptide and its preparation method
CNA031204007A CN1532284A (en) 2002-11-12 2003-03-19 Novel antibiotic polypeptide and its preparing method

Publications (1)

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CN1532284A true CN1532284A (en) 2004-09-29

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006109192A2 (en) * 2005-01-21 2006-10-19 West China Hospital, Sichuan University Antifungal bifunctional molecules, methods of construction and methods of treating fungal infection therewith
WO2007083175A1 (en) * 2006-01-17 2007-07-26 West China Hospital, Sichuan University Antiviral bifunctional molecules, methods of construction and methods of treating virus-induced cancer therewith
CN101643501B (en) * 2008-11-07 2012-06-20 畿晋庆三联（北京）生物技术有限公司 Novel antibiotic and nucleotide sequence, preparation method and application thereof
CN107365363A (en) * 2017-09-20 2017-11-21 苏州大学 Micromolecule polypeptide and its application

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006109192A2 (en) * 2005-01-21 2006-10-19 West China Hospital, Sichuan University Antifungal bifunctional molecules, methods of construction and methods of treating fungal infection therewith
WO2006109192A3 (en) * 2005-01-21 2007-06-14 West China Hospital Sichuan Univ Antifungal bifunctional molecules, methods of construction and methods of treating fungal infection therewith
CN1330758C (en) * 2005-01-21 2007-08-08 四川大学 Antifungal polypeptide and its preparation method
WO2007083175A1 (en) * 2006-01-17 2007-07-26 West China Hospital, Sichuan University Antiviral bifunctional molecules, methods of construction and methods of treating virus-induced cancer therewith
CN101643501B (en) * 2008-11-07 2012-06-20 畿晋庆三联（北京）生物技术有限公司 Novel antibiotic and nucleotide sequence, preparation method and application thereof
CN107365363A (en) * 2017-09-20 2017-11-21 苏州大学 Micromolecule polypeptide and its application
CN107365363B (en) * 2017-09-20 2020-08-04 苏州大学 Small molecule polypeptide and application thereof

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