CN101302526A - Recombinant soluble streptococcus hemolyticus haemolysin O gene, recombinant protein and preparation thereof - Google Patents
Recombinant soluble streptococcus hemolyticus haemolysin O gene, recombinant protein and preparation thereof Download PDFInfo
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- CN101302526A CN101302526A CNA2008100454121A CN200810045412A CN101302526A CN 101302526 A CN101302526 A CN 101302526A CN A2008100454121 A CNA2008100454121 A CN A2008100454121A CN 200810045412 A CN200810045412 A CN 200810045412A CN 101302526 A CN101302526 A CN 101302526A
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Abstract
The invention relates to a recombinant protein for providing a soluble hemolysis streptolysin O gene and the gene expression, and a method for making the same. The method comprises the steps of: cloning from the streptococcus hemolyticus gene group by using the gene engineering technology to obtain a truncated SLO gene; connecting to an expression vector and transferring into host cell expression to purify a large amount of stable r-s SLO proteins, thereby overcoming various shortcomings of the protein from natural extraction and being favorable for standardization of making various reagents by using the protein as the raw material. The whole-length SLO protein is the protein of about 60 KD containing 571 AA, and the end N is provided with a signal peptide which is used for guiding the mature protein out of the cell. A research shows that the end N containing 77 AA provided with the signal peptide has a hazard function to the host cell, wherein the hazard function is more than a lyticase function which is generated by the latter AA of between 78 and 571.
Description
Technical field
The invention belongs to biological technical field, particularly, relate to recombinant protein of a kind of recombinant soluble streptococcus hemolyticus haemolysin O gene, this genetic expression and preparation method thereof.
Background technology
Streptococcus hemolyticus bacteriolysin O is that multiple A type, G type and C type Streptococcus hemolyticus all can have molten cell characteristic protein by excretory.The people infects the specific antibody that body behind the Streptococcus hemolyticus can produce the anti-hemolysis streptolysin O, and the concentration that detects this specific antibody has very important significance to clinical diagnosis; Streptococcus hemolyticus haemolysin O also is used to comprise the field, a plurality of forward position of cancer therapy in recent years widely because it has the biologic activity of punching on the cytolemma of cholesterol containing simultaneously.The purification hemolysin O has and much is difficult to the shortcoming that overcomes from natural Streptococcus hemolyticus, and for example: yield is very low; Difference between batch is very big; Be difficult to suitability for industrialized production; There is Biosafety hidden danger etc.
Summary of the invention
Technical problem to be solved by this invention provides recombinant protein of a kind of soluble streptococcus hemolyticus haemolysin O gene, this genetic expression and preparation method thereof.
The present invention solves the problems of the technologies described above the technical scheme that is adopted: the soluble streptococcus hemolyticus haemolysin O gene of nucleotide sequence shown in a kind of SEQ of having IDNO:1.
The present invention uses genetic engineering technique to clone the SLO gene that obtains brachymemma from the Streptococcus hemolyticus genome, be connected to and be transformed into a large amount of stable r-sSLO albumen of host cell expression purifying on the expression vector again, overcome the various shortcomings of natural extract, helping with this albumen is the stdn of raw material preparing all ingredients.Total length SLO albumen is the albumen that contains about the 60KD of 571 AA, and the N end has signal peptide to be used for maturation protein is guided to the extracellular.Studies show that 77 AA of N end that comprise signal peptide to the toxic effect of host cell, its toxic action is greater than 78-571 molten cytosis that AA produced thereafter.(wherein, SLO is a soluble streptococcus hemolyticus haemolysin O, and r-sSLO is a recombinant soluble streptococcus hemolyticus haemolysin O, and AA is an amino acid)
A kind of recombinant soluble streptococcus hemolyticus haemolysin O albumen with aminoacid sequence shown in the SEQ ID NO:2.
Reorganization SLO albumen provided by the invention is about to 77 aminoacid deletion sudden changes of the proteic N end of SLO, removes signal peptide and the bigger amino acid segment of toxicity, obtains having with the mutant of the identical dissolved cell activity of natural SLO albumen and with its called after r-sSLO albumen.
The proteic method of recombinant soluble streptococcus hemolyticus haemolysin O shown in a kind of preparation SEQ ID NO:2 comprises the steps: (1) isolation of genomic DNA from Streptococcus hemolyticus, obtains recombinant soluble streptococcus hemolyticus haemolysin O cDNA with PCR method; (2) cDNA sequence and the expression plasmid reorganization that obtains with step (1), recon are determined the sequence of recombinant plasmid and read frame all correct through the dna sequencing checking; (3) expression vector with step (2) transforms expression host cell; (4) cultivate host cell and from substratum, reclaiming and purification of Recombinant soluble streptococcus hemolyticus haemolysin O albumen.
The present invention prepares the proteic method of r-sSLO, comprises the r-sSLOcDNA gene for preparing biologically active, be cloned in the expression vector, and transformed host cell, the inducing culture host cell, and purifying prepares r-sSLO.The present invention is isolation of genomic DNA from Streptococcus hemolyticus, obtains the soluble streptococcus hemolyticus haemolysin O cDNA of deletion mutantion with PCR method, the cDNA sequence of acquisition and expression plasmid reorganization, and recon is through the dna sequencing checking, and base is with to read frame all correct.The invention is not restricted to specific expression vector, in a preferred embodiment, the present invention uses prokaryotic expression carrier, for example: PET32a (+).Above-mentioned recombinant expression vector imports host cell according to a conventional method.The invention is not restricted to any host cell, as long as can expressing said gene.In a preferred embodiment, the present invention uses intestinal bacteria Rosseta gamiII (DE3), and Rosseta gami II (DE3) purchases the company in Merck, is MerK company oneself to the name of intestinal bacteria product that specific function is arranged.Expression product of the present invention is present in the super supernatant after broken of thalline, but obtains desired product with nickel post single step purification.
To sum up, the invention has the beneficial effects as follows: the present invention uses genetic engineering technique to clone the SLO gene that obtains brachymemma from the Streptococcus hemolyticus genome, be connected to and be transformed into a large amount of stable r-sSLO albumen of host cell expression purifying on the expression vector again, overcome the various shortcomings of natural extract, helping with this albumen is the stdn of raw material preparing all ingredients.
Embodiment
For reaching the object of the invention, the present invention prepares r-sSLO albumen and comprises the steps:
(1) clone r-sSLO gene, construction expression plasmid r-sSLO-PET32a (+)
Design primer according to SLO aminopeptidase gene acid order, this gene of pcr amplification also is connected with the pMD18-T carrier, and transformed into escherichia coli DH5 a, PCR identify that mono-clonal determines possible positive plasmid, extract the evaluation of plasmid double digestion again.The positive plasmid that process dna sequencing proof base and open reading frame are correct cuts out with the PET32a (+) of same double digestion with suitable enzyme and is connected, and structure prokaryotic expression plasmid r-sSLO-PET32a (+) transforms expressive host bacterium Rosseta gamiII.Double digestion is identified positive host bacterium.Above-mentioned experiment is this laboratory with plasmid vector and preserves, and expresses bacterium Rosseta gami II and purchases the company in Merck, and restriction enzyme is purchased the company in Dalian Takara.
(2) screen the positive expression bacterial strain that efficiently expresses
Inoculate among above-mentioned positive strain of 10 strains and the 5mlLB+amp, 37 ℃, the 220rpm overnight incubation was inoculated in the fresh LB+amp substratum 37 ℃ by 1% inoculum size on 1st, 220rpm cultivated 4-5 hour, get 1ml bacterium liquid and do tropina contrast under the inductive condition not, remaining bacterium liquid adds the IPTG of final concentration 0.5mM, 25 ℃, the 170rpm inducing culture spends the night, and gets 1ml bacterium liquid and contrasts as the tropina after inducing.
The centrifugal supernatant that goes of bacterium bacterium liquid before and after inducing, thalline uses the TE damping fluid of 100ul resuspended respectively, boil 5min after, the SDS-PAGE electrophoresis.Coomassie brilliant blue dyeing, BandScan analytical electrophoresis band.Bacterium after inducing has a protein band that concentrates at the molecular weight place of prediction, and the bacterium before inducing does not have protein band at identical molecular weight place.Account for more than 50% of total protein concentration through this target protein of BandScan software analysis.Selecting the high bacterial strain of expression level is engineering strain.
After the bacterium ultrasonication after inducing on the centrifuging and taking cleer and peaceful precipitation do not carry out the SDS-PAGE electrophoretic analysis, the result shows that recombinant protein is to be present in the supernatant with soluble form.
(3) purifying of the enlarged culturing of engineering bacteria and target protein
According to a small amount of culture condition enlarged culturing amount, 1% glycerol stock is inoculated in the 20mlLB+amp substratum 37 ℃, and 220rpm incubated overnight, second day 2% inoculum size are inoculated in the 1LLB+amp substratum 37 ℃, and 220rpm cultivates 4-5h to OD
600To 0.5-1.0, add IPTG to final concentration be 0.5mM, 25 ℃, 170rpm inducing culture (12-16h) the centrifugal collection thalline that spends the night is pressed the 1G thalline and is added 10ml Loading Buffer (20Mm PBS, 0.5MNaCl, the 30mM imidazoles, ratio PH7.4) adds Loading Buffer, the ultrasonication recentrifuge is got supernatant, prepares against purifying with the membrane filtration of 0.45uM.
HisTRAP H.P (GE company) post, with 10 times of column volume Loading Buffer balance pillars, the direct upper prop of above-mentioned filtration supernatant is washed post to OD with Loading Buffer behind the last sample
280Absorption value is to baseline, with Elusion Buffer (20Mm PBS, 0.5M NaCl, the 300mM imidazoles, PH7.4) wash-out is collected elution peak, carries out the SDS-PAGE purity assay, the result shows that purity reaches more than 60%, protein concentration 2-3mg/ml.The 6L nutrient solution can obtain 140-200mg albumen.
(4) Elisa and Western identify recombinant protein antigen
With distinguishing wrapper sheet according to the Elisa standard operating procedure after anti-" O " feminine gender and 1: 50 times of dilution of positive definite value serum, develop the color respectively with after the r-sSLO albumen test, the result shows that r-sSLO can obviously distinguish anti-" O " positive and negative serum.In the Western experiment, one anti-ly is anti-" O " feminine gender, 1: 500 times of diluent of positive serum, and two anti-ly are HRP mark mouse-anti human IgG1: 4000 times of diluents, the result is identical with Elisa, has shown that the r-sSLO reaction sensitivity is high.
Sequence table
＜110〉Maike Tech Co., Ltd., Sichuan Prov.
＜120〉recombinant soluble streptococcus hemolyticus haemolysin O gene, recombinant protein and preparation method thereof
＜130〉recombinant soluble streptococcus hemolyticus haemolysin O gene, recombinant protein and preparation method thereof
<160>2
<170>PatentIn?version?3.3
<210>1
<211>1440
<212>DNA
＜213〉Streptococcus hemolyticus Streptococcus pyogenes
<400>1
cttgctccca?aagaaatgcc?actagaatct?gcagaaaaag?aagaaaaaaa?gtcagaagac 60
aaaaaaaaga?gcgaagaaga?tcacactgaa?gaaatcaatg?acaagattta?ttcactaaat 120
tataatgagc?ttgaagtact?tgctaaaaat?ggtgaaacca?ttgaaaattt?tgttcctaaa 180
gaaggcgtta?agaaagctga?taaatttatt?gtcattgaaa?gaaagaaaaa?aaatatcaac 240
actacaccag?tcgatatttc?cattattgac?tctgtcactg?ataggaccta?tccagcagcc 300
cttcagctgg?ctaataaagg?ttttaccgaa?aacaaaccag?acgcggtagt?caccaagcga 360
aacccacaaa?aaatccatat?tgatttacca?ggtatgggag?acaaagcaac?ggttgaggtc 420
aatgacccta?cctatgccaa?tgtttcaaca?gctattgata?atcttgttaa?ccaatggcat 480
gataattatt?ctggtggtaa?tacgcttcct?gccagaacac?aatatactga?atcaatggta 540
tattctaagt?cacagattga?agcagctcta?aatgttaata?gcaaaatctt?agatggtact 600
ttaggcattg?atttcaagtc?gatttcaaaa?ggtgaaaaga?aggtgatgat?tgcagcatac 660
aagcaaattt?tttacaccgt?atcagcaaac?cttcctaata?atcctgcgga?tgtgtttgat 720
aaatcagtga?cctttaaaga?tttgcaacga?aaaggtgtca?gcaatgaagc?tccgccactc 780
tttgtgagta?acgtagccta?tggtcgaact?gtttttgtca?aactagaaac?aagttctaaa 840
agtaatgatg?ttgaagcggc?ctttagtgca?gctctaaaag?gaacagatgt?taaaacgaat 900
ggaaaatact?ctgatatctt?agaaaatagc?tcatttacag?ctgtcgtttt?aggaggagat 960
gctgcagagc?acaataaggt?ggtcacaaaa?gactttgatg?ttattagaaa?cgttatcaaa 1020
gacaatgcta?ccttcagtag?aaaaaaccca?gcttatccta?tttcatacac?cagtgttttc 1080
cttaaaaata?ataaaattgc?gggtgtcaat?aacagaactg?aatacgttga?aacaacatct 1140
accgagtaca?ctagtggaaa?aattaacctg?tctcatcaag?gcgcgtatgt?tgctcaatat 1200
gaaatccttt?gggatgaaat?caattatgat?gacaaaggaa?aagaagtgat?tacaaaacga 1260
cgttgggaca?acaactggta?tagtaagaca?tcaccattta?gcacagttat?cccactagga 1320
gctaattcac?gaaatatccg?tatcatggct?agagagtgca?ccggcttagc?ttgggaatgg 1380
tggcgaaaag?tgatcgacga?aagagatgtg?aaactatcta?aagaaatcaa?tgtcaacatc 1440
<210>2
<211>494
<212>PRT
＜213〉Streptococcus hemolyticus Streptococcus pyogenes
<400>2
Leu?Ala?Pro?Lys?Glu?Met?Pro?Leu?Glu?Ser?Ala?Glu?Lys?Glu?Glu?Lys
1 5 10 15
Lys?Ser?Glu?Asp?Lys?Lys?Lys?Ser?Glu?Glu?Asp?His?Thr?Glu?Glu?Ile
20 25 30
Asn?Asp?Lys?Ile?Tyr?Ser?Leu?Asn?Tyr?Asn?Glu?Leu?Glu?Val?Leu?Ala
35 40 45
Lys?Asn?Gly?Glu?Thr?Ile?Glu?Asn?Phe?Val?Pro?Lys?Glu?Gly?Val?Lys
50 55 60
Lys?Ala?Asp?Lys?Phe?Ile?Val?Ile?Glu?Arg?Lys?Lys?Lys?Asn?Ile?Asn
65 70 75 80
Thr?Thr?Pro?Val?Asp?Ile?Ser?Ile?Ile?Asp?Ser?Val?Thr?Asp?Arg?Thr
85 90 95
Tyr?Pro?Ala?Ala?Leu?Gln?Leu?Ala?Asn?Lys?Gly?Phe?Thr?Glu?Asn?Lys
100 105 110
Pro?Asp?Ala?Val?Val?Thr?Lys?Arg?Asn?Pro?Gln?Lys?Ile?His?Ile?Asp
115 120 125
Leu?Pro?Gly?Met?Gly?Asp?Lys?Ala?Thr?Val?Glu?Val?Asn?Asp?Pro?Thr
130 135 140
Tyr?Ala?Asn?Val?Ser?Thr?Ala?Ile?Asp?Asn?Leu?Val?Asn?Gln?Trp?His
145 150 155 160
Asp?Asn?Tyr?Ser?Gly?Gly?Asn?Thr?Leu?Pro?Ala?Arg?Thr?Gln?Tyr?Thr
165 170 175
Glu?Ser?Met?Val?Tyr?Ser?Lys?Ser?Gln?Ile?Glu?Ala?Ala?Leu?Asn?Val
180 185 190
Asn?Ser?Lys?Ile?Leu?Asp?Gly?Thr?Leu?Gly?Ile?Asp?Phe?Lys?Ser?Ile
195 200 205
Ser?Lys?Gly?Glu?Lys?Lys?Val?Met?Ile?Ala?Ala?Tyr?Lys?Gln?Ile?Phe
210 215 220
Tyr?Thr?Val?Ser?Ala?Asn?Leu?Pro?Asn?Asn?Pro?Ala?Asp?Val?Phe?Asp
225 230 235 240
Lys?Ser?Val?Thr?Phe?Lys?Asp?Leu?Gln?Arg?Lys?Gly?Val?Ser?Asn?Glu
245 250 255
Ala?Pro?Pro?Leu?Phe?Val?Ser?Asn?Val?Ala?Tyr?Gly?Arg?Thr?Val?Phe
260 265 270
Val?Lys?Leu?Glu?Thr?Ser?Ser?Lys?Ser?Asn?Asp?Val?Glu?Ala?Ala?Phe
275 280 285
Ser?Ala?Ala?Leu?Lys?Gly?Thr?Asp?Val?Lys?Thr?Asn?Gly?Lys?Tyr?Ser
290 295 300
Asp?Ile?Leu?Glu?Asn?Ser?Ser?Phe?Thr?Ala?Val?Val?Leu?Gly?Gly?Asp
305 310 315 320
Ala?Ala?Glu?His?Asn?Lys?Val?Val?Thr?Lys?Asp?Phe?Asp?Val?Ile?Arg
325 330 335
Asn?Val?Ile?Lys?Asp?Asn?Ala?Thr?Phe?Ser?Arg?Lys?Asn?Pro?Ala?Tyr
340 345 350
Pro?Ile?Ser?Tyr?Thr?Ser?Val?Phe?Leu?Lys?Asn?Asn?Lys?Ile?Ala?Gly
355 360 365
Val?Asn?Asn?Arg?Thr?Glu?Tyr?Val?Glu?Thr?Thr?Ser?Thr?Glu?Tyr?Thr
370 375 380
Ser?Gly?Lys?Ile?Asn?Leu?Ser?His?Gln?Gly?Ala?Tyr?Val?Ala?Gln?Tyr
385 390 395 400
Glu?Ile?Leu?Trp?Asp?Glu?Ile?Asn?Tyr?Asp?Asp?Lys?Gly?Lys?Glu?Val
405 410 415
Ile?Thr?Lys?Arg?Arg?Trp?Asp?Asn?Asn?Trp?Tyr?Ser?Lys?Thr?Ser?Pro
420 425 430
Phe?Ser?Thr?Val?Ile?Pro?Leu?Gly?Ala?Asn?Ser?Arg?Asn?Ile?Arg?Ile
435 440 445
Met?Ala?Arg?Glu?Cys?Thr?Gly?Leu?Ala?Trp?Glu?Trp?Trp?Arg?Lys?Val
450 455 460
Ile?Asp?Glu?Arg?Asp?Val?Lys?Leu?Ser?Lys?Glu?Ile?Asn?Val?Asn?Ile
465 470 475 480
Ser?Gly?Ser?Thr?Leu?Ser?Pro?Tyr?Gly?Ser?Ile?Thr?Tyr?Lys
485 490
Claims (10)
1, a kind of recombinant soluble streptococcus hemolyticus haemolysin O gene with nucleotide sequence shown in the SEQ ID NO:1.
2, a kind of recombinant soluble streptococcus hemolyticus haemolysin O albumen with aminoacid sequence shown in the SEQ ID NO:2.
3, the proteic method of recombinant soluble streptococcus hemolyticus haemolysin O shown in a kind of preparation SEQ ID NO:2 is characterized in that, comprises the steps:
(1) isolation of genomic DNA from Streptococcus hemolyticus obtains recombinant soluble streptococcus hemolyticus haemolysin O cDNA with PCR method;
(2) cDNA sequence and the expression plasmid reorganization that obtains with step (1), recon are determined the sequence of recombinant plasmid and read frame all correct through the dna sequencing checking;
(3) expression vector with step (2) transforms expression host cell;
(4) cultivate host cell and from substratum, reclaiming and purification of Recombinant soluble streptococcus hemolyticus haemolysin O albumen.
4, the proteic method of recombinant soluble streptococcus hemolyticus haemolysin O shown in the preparation SEQ ID NO:2 according to claim 3, it is characterized in that described expression vector is can recombinate with the proteic gene order of recombinant soluble streptococcus hemolyticus haemolysin O to form the expression vector of expression plasmid.
5, the proteic method of recombinant soluble streptococcus hemolyticus haemolysin O shown in the preparation SEQ ID NO:2 according to claim 4 is characterized in that described carrier is a prokaryotic expression carrier.
6, the proteic method of recombinant soluble streptococcus hemolyticus haemolysin O shown in the preparation SEQ ID NO:2 according to claim 5 is characterized in that described prokaryotic expression carrier is PET32a (+).
7, the proteic method of recombinant soluble streptococcus hemolyticus haemolysin O shown in the preparation SEQ ID NO:2 according to claim 3 is characterized in that, described expression host cell is for expressing the expression host cell of described expression plasmid.
8, the proteic method of recombinant soluble streptococcus hemolyticus haemolysin O shown in the preparation SEQ ID NO:2 according to claim 3 is characterized in that described expression host cell is intestinal bacteria Rosseta gami II (DE3).
9, the proteic method of recombinant soluble streptococcus hemolyticus haemolysin O shown in the preparation SEQ ID NO:2 according to claim 3, it is characterized in that, described substratum adopts and contains microbiotic basic medium amplification cultivation host cell, parameter is: 35 ℃-37 ℃ of culture temperature, incubation time 4～5h; 22 ℃～25 ℃ of inducing temperatures, induction time 14～18h; Inductor IPTG final concentration is 0.2～0.5mM.
10, the proteic method of recombinant soluble streptococcus hemolyticus haemolysin O shown in the preparation SEQ ID NO:2 according to claim 3, it is characterized in that, described recombinant soluble streptococcus hemolyticus haemolysin O albumen is got supernatant and is obtained through nickel post single step purification by centrifugal collection thalline behind the engineering bacteria inducing culture after the ultrasonication.
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| CNA2008100454121A CN101302526A (en) | 2008-06-27 | 2008-06-27 | Recombinant soluble streptococcus hemolyticus haemolysin O gene, recombinant protein and preparation thereof |
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| CNA2008100454121A CN101302526A (en) | 2008-06-27 | 2008-06-27 | Recombinant soluble streptococcus hemolyticus haemolysin O gene, recombinant protein and preparation thereof |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102010870A (en) * | 2010-06-28 | 2011-04-13 | 盐城师范学院 | High-expression streptolysin O (SLO) gene as well as secreting expression vector and application thereof |
| CN102964435A (en) * | 2011-08-31 | 2013-03-13 | 北京利德曼生化股份有限公司研发中心 | Truncated-form streptococcus hemolyticus bacteriolysin O and detection kit using same |
| CN104098659A (en) * | 2014-07-22 | 2014-10-15 | 桂林英美特生物技术有限公司 | Recombinant SLO (Streptolysin O) protein and preparation method thereof |
| CN107167586A (en) * | 2017-07-06 | 2017-09-15 | 四川新健康成生物股份有限公司 | A kind of antistreptolysin O (ASO) detection kit and preparation method thereof |
| CN113880957A (en) * | 2021-11-04 | 2022-01-04 | 上海捷门生物技术有限公司 | Streptolysin O fusion protein |
| WO2022158373A1 (en) * | 2021-01-19 | 2022-07-28 | デンカ株式会社 | Protein, hemolytic streptococcus vaccine, dna, vector, transformant, method for producing protein, baculovirus, agrobacterium, latex particle, kit, and method for measuring anti-streptolysin o antibody |
| CN114854771A (en) * | 2022-05-11 | 2022-08-05 | 武汉爱博泰克生物科技有限公司 | Recombinant expression of thiol-activated cytolysin proteins |
| CN115947805A (en) * | 2022-10-20 | 2023-04-11 | 南京立顶医疗科技有限公司 | Preparation method of streptolysin O protein with double high characteristics |
-
2008
- 2008-06-27 CN CNA2008100454121A patent/CN101302526A/en active Pending
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102010870A (en) * | 2010-06-28 | 2011-04-13 | 盐城师范学院 | High-expression streptolysin O (SLO) gene as well as secreting expression vector and application thereof |
| CN102010870B (en) * | 2010-06-28 | 2013-06-12 | 盐城师范学院 | High-expression streptolysin O (SLO) gene as well as secreting expression vector and application thereof |
| CN102964435A (en) * | 2011-08-31 | 2013-03-13 | 北京利德曼生化股份有限公司研发中心 | Truncated-form streptococcus hemolyticus bacteriolysin O and detection kit using same |
| CN102964435B (en) * | 2011-08-31 | 2014-12-03 | 北京利德曼生化股份有限公司研发中心 | Truncated-form streptococcus hemolyticus bacteriolysin O and detection kit using same |
| CN104098659A (en) * | 2014-07-22 | 2014-10-15 | 桂林英美特生物技术有限公司 | Recombinant SLO (Streptolysin O) protein and preparation method thereof |
| CN107167586A (en) * | 2017-07-06 | 2017-09-15 | 四川新健康成生物股份有限公司 | A kind of antistreptolysin O (ASO) detection kit and preparation method thereof |
| WO2022158373A1 (en) * | 2021-01-19 | 2022-07-28 | デンカ株式会社 | Protein, hemolytic streptococcus vaccine, dna, vector, transformant, method for producing protein, baculovirus, agrobacterium, latex particle, kit, and method for measuring anti-streptolysin o antibody |
| CN113880957A (en) * | 2021-11-04 | 2022-01-04 | 上海捷门生物技术有限公司 | Streptolysin O fusion protein |
| CN114854771A (en) * | 2022-05-11 | 2022-08-05 | 武汉爱博泰克生物科技有限公司 | Recombinant expression of thiol-activated cytolysin proteins |
| CN115947805A (en) * | 2022-10-20 | 2023-04-11 | 南京立顶医疗科技有限公司 | Preparation method of streptolysin O protein with double high characteristics |
| CN115947805B (en) * | 2022-10-20 | 2023-10-13 | 南京立顶医疗科技有限公司 | Preparation method of streptolysin O protein with double high characteristics |
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