猪带绦虫重组Bb(pGEX-TSO45W-4B-TSOL18)疫苗的构建及鉴定  

Construction and identification of a recombinant Bb(pGEX-TSO45W-4B-TSOL18)vaccine of Taenia solium

作　　者：周必英[1] 刘美辰[1] 贺莉芳[1] 

机构地区：[1]遵义医学院寄生虫学教研室,贵州遵义563003

出　　处：《中国病原生物学杂志》2014年第4期289-292,298,共5页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.81160206)

摘　　要：目的构建和鉴定猪带绦虫重组双歧杆菌(Bb)(pGEX-TSO45W-4B-TSOL18)疫苗。方法用疏水甘氨酸接头连接TSO45W-4B和TSOL18编码基因,通过全基因合成方法合成猪带绦虫TSO45W-4B-TSOL18融合基因。将该融合基因定向克隆到大肠埃希菌-双歧杆菌穿梭表达载体pGEX-1λT中,构建重组质粒pGEX-TSO45W-4B-TSOL18。电穿孔法将该质粒导入Bb,构建猪带绦虫重组Bb(pGEX-TSO45W-4B-TSOL18)疫苗,从具有氨苄青霉素抗性的rBb中抽提质粒进行酶切、PCR和测序鉴定。结果全基因合成789bp的TSO45W-4B-TSOL18融合基因片段。从具有氨苄青霉素抗性的rBb中抽提质粒pGEX-TSO45W-4B-TSOL18,经BamHⅠ和EcoRⅠ双酶切得到4 944bp的pGEX-1λT载体片段和789bp的TSO45W-4B-TSOL18融合基因片段,与预期结果相符;以该重组质粒为模板进行PCR扩增得到789bp的TSO45W-4B-TSOL18融合基因片段,经测序基因片段为789bp,与预期大小相符。结论成功构建了猪带绦虫重组Bb(pGEX-TSO45W-4B-TSOL18)疫苗,为该疫苗的表达及免疫原性研究奠定了基础。Objectives To construct and identify a recombinant Bb-TSO45W-4B-TSOL18vaccine of Taenia solium. Methods Genes encoding TSO45W-4Band TSOL18were fused with a（Gly4Ser）3hydrophobic linker.The TSO45W-4BTSOL18fusion gene was synthesized and cloned into the Escherichia coli-Bifidobacteria shuttle vector pGEX-1λT to construct the recombinant plasmid pGEX-TSO45W-4B-TSOL18.The recombinant plasmid was electroporated into Bifidobacteria（Bb）to construct a recombinant Bb（pGEX-TSO45W-4B-TSOL18）vaccine.The plasmid was extracted from ampicillin-resistant recombinant Bifidobacteriaand identified using restriction endonuclease digestion,PCR,and sequencing. Results A TSO45W-4B-TSOL18fusion gene of 789bp was synthesized.The products of restriction endonuclease digestion were the same as would be expected from digestion using BamHⅠ and EcoRⅠ,including apGEX-1λT vector fragment of 4 944bp and a TSO45W-4B-TSOL18fusion gene fragment of 789bp.A TSO45W-4B-TSOL18fusion gene fragment of 789bp was amplified with PCR using the recombinant plasmid pGEX-TSO45W-4B-TSOL18extracted from the recombinant Bifidobacteriavaccine as a template.DNA sequencing indicated that the fusion gene TSO45W-4BTSOL18was 789bp in length,which was consistent with the expected result. Conclusion A recombinant Bb（pGEXTSO45W-4B-TSOL18）vaccine of Taenia solium was successfully constructed,and this lays the foundation for study of the expression and immunogenicity of the vaccine.

关 键 词：猪带绦虫 重组Bb(pGEX-TSO45W-4B-TSOL18)疫苗 构建 鉴定 

分 类 号：R383.32[医药卫生—医学寄生虫学]