Meng-Meng Yin, Qiu-Dong Su, Min-Zhuo Gyo, Yi-Na Cun, Yuan-Qian Pu, Zhi-Yuan Jia, Jing-Ran Yang, Yang Tang, Guo-Yang Liao, Yao Yi, Sheng-Li Bi, Wei-Dong Li
{"title":"b型流感嗜血杆菌蛋白D的原核可溶性表达","authors":"Meng-Meng Yin, Qiu-Dong Su, Min-Zhuo Gyo, Yi-Na Cun, Yuan-Qian Pu, Zhi-Yuan Jia, Jing-Ran Yang, Yang Tang, Guo-Yang Liao, Yao Yi, Sheng-Li Bi, Wei-Dong Li","doi":"","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>To express the recombinant D protein in prokaryotic expression system solubly and make preparation for producing D-carrier conjugate vaccine next step.</p><p><strong>Methods: </strong>The hpd gene fragment removed of signal peptide from genomic DNA of Hib CMCC was inserted into pET43. 1a. The recombinant plasmid was transformed to competent E. coli BL21 (DE3) for expression under induction of IPTG. The expressed recombination protein was precipitated with ammonium sulfate, purified by DEAE anion exchange column chromatography and identified for reactogenicity by Western Blot.</p><p><strong>Results: </strong>The expressed recombination protein, in a soluble form, constained about 50% of total somatic protein and showed specific reaction with the HIB antisera after preliminary purification.</p><p><strong>Conclusion: </strong>The D protein recombined expression plasmid was constructed successfully and expressed D protein in prokaryotic cells in a solube form.</p>","PeriodicalId":70973,"journal":{"name":"中华实验和临床病毒学杂志","volume":"27 2","pages":"89-91"},"PeriodicalIF":0.0000,"publicationDate":"2013-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"[Prokaryotic soluble expression of protein D of Haemophilus influenzae type b].\",\"authors\":\"Meng-Meng Yin, Qiu-Dong Su, Min-Zhuo Gyo, Yi-Na Cun, Yuan-Qian Pu, Zhi-Yuan Jia, Jing-Ran Yang, Yang Tang, Guo-Yang Liao, Yao Yi, Sheng-Li Bi, Wei-Dong Li\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Objective: </strong>To express the recombinant D protein in prokaryotic expression system solubly and make preparation for producing D-carrier conjugate vaccine next step.</p><p><strong>Methods: </strong>The hpd gene fragment removed of signal peptide from genomic DNA of Hib CMCC was inserted into pET43. 1a. The recombinant plasmid was transformed to competent E. coli BL21 (DE3) for expression under induction of IPTG. The expressed recombination protein was precipitated with ammonium sulfate, purified by DEAE anion exchange column chromatography and identified for reactogenicity by Western Blot.</p><p><strong>Results: </strong>The expressed recombination protein, in a soluble form, constained about 50% of total somatic protein and showed specific reaction with the HIB antisera after preliminary purification.</p><p><strong>Conclusion: </strong>The D protein recombined expression plasmid was constructed successfully and expressed D protein in prokaryotic cells in a solube form.</p>\",\"PeriodicalId\":70973,\"journal\":{\"name\":\"中华实验和临床病毒学杂志\",\"volume\":\"27 2\",\"pages\":\"89-91\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2013-04-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"中华实验和临床病毒学杂志\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"中华实验和临床病毒学杂志","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
[Prokaryotic soluble expression of protein D of Haemophilus influenzae type b].
Objective: To express the recombinant D protein in prokaryotic expression system solubly and make preparation for producing D-carrier conjugate vaccine next step.
Methods: The hpd gene fragment removed of signal peptide from genomic DNA of Hib CMCC was inserted into pET43. 1a. The recombinant plasmid was transformed to competent E. coli BL21 (DE3) for expression under induction of IPTG. The expressed recombination protein was precipitated with ammonium sulfate, purified by DEAE anion exchange column chromatography and identified for reactogenicity by Western Blot.
Results: The expressed recombination protein, in a soluble form, constained about 50% of total somatic protein and showed specific reaction with the HIB antisera after preliminary purification.
Conclusion: The D protein recombined expression plasmid was constructed successfully and expressed D protein in prokaryotic cells in a solube form.
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