| Construtcion of Neisseria Gonorrhoeae Porin B Plasmid Recombinant and Its Expression in E.coli |
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| 作者姓名： | 宋启发 廖芳 叶嗣颖 崔斌 熊萍 |
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| 作者单位： | Department of Biopathogen，School of Basic Medical Sciences，Tong ji Medical College，Huazhong University of Science and Technology，Department of Biopathogen，School of Basic Medical Sciences，Tong ji Medical College，Huazhong University of Science and Technology，Department of Biopathogen，School of Basic Medical Sciences，Tong ji Medical College，Huazhong University of Science and Technology，Department of Biopathogen，School of Basic Medical Sciences，Tong ji Medical College，Huazhong University of Science and Technology，Department of Biopathogen，School of Basic Medical Sciences，Tong ji Medical College，Huazhong University of Science and Technology Wuhan 430030，China，Wuhan 430030，China，Wuhan 430030，China，Wuhan 430030，China，Wuhan 430030，China |
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| 摘 要： | Neisseria gonorrhoeaeis a common pathogenicmicroorganism which causes sex transmitted dis-ease . The porins ,the predominant proteins on thesurface of pathogenicNeisseria,form a family ofstructurally related proteins whose physiologicalrole is to allownutrients access into the cell . Theyalso play ani mportant role in pathogenesis ,gener-atingi mmunological specificity andinteracting withthe host cell .Neisseria gonorrhoeaeexpresses oneof the two alternative classes of porins PIA andPIB1].…
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Construtcion of Neisseria Gonorrhoeae Porin B Plasmid Recombinant and Its Expression in E.coli |
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| SONG Qifa,Liao Fang,YE Siying,CUI Bing,XIONG Ping.Construtcion of Neisseria Gonorrhoeae Porin B Plasmid Recombinant and Its Expression in E.coli[J].Journal of Zuazhong University of Science and Technology: Medical Edition,2005,25(3). |
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| Authors: | SONG Qifa Liao Fang YE Siying CUI Bing XIONG Ping |
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| Institution: | Department of Biopathogen, School of Basic Medical Sciences, Tong ji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China |
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| Abstract: | A prokaryotic expression recombinant plasmid pET-PIB to express porin B (PIB) of Neisseria gonorrhoeae in E.coli DE_3 was constructed in order to provide a basis of research in detection, prophylactic and therapeutic vaccine against the pathogen infection. The gene encoding PIB was amplified by PCR from Neisseria gonorrhoeae and cloned into prokaryotic expression plasmid pET-28a(+) to construct a pET-PIB recombinant, which was verified by restriction endonuclease and DNA sequencing. Protein PIB was expressed in E.coli DE_3 induced with IPTG. The antigenicity of the expressed protein was evaluated by indirect ELISA. Rabbits were immunized with the protein and serum was collected after immunization. To assess the immunogenicity of the protein, the titer of serum to protein PIB was determined by ELISA. DNA sequence analysis showed that the nucleic acid sequence of PIB gene was 99.28 % of homology compared with that (NGPIB18) published in GenBank. A 41 kD fused protein was detected by SDS-PAGE and was proven to have reactivity with anti-PIB polyclonal antibody from mouse. A polyclonal antibody to PIB of 1:4000 titer determined by indirect ELISA was obtained from rabbit immunized with the purified product. Recombinant plasmid encoding PIB of Neisseria gonorrhoeae was constructed. Protein PIB with antigenicity and immunogenicity was successfully expressed. |
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| Keywords: | Neisseria gonorrhoeae porin B prokaryotic expression |
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