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Position: Home > Articles > Cloning and expression of toxB gene in segments from enterohemorrhagic Escherichia coli O157 ∶ H7 Jiangsu Journal of Agricultural Sciences 2014 (6) 1392-1395

肠出血性大肠杆菌O157∶H7toxB基因的分片段克隆和表达

作  者:
张雪寒;栾晓婷;何孔旺;倪艳秀;周俊明
单  位:
江苏省农业科学院兽医研究所/农业部兽用生物制品工程技术重点实验室/国家兽用生物制品工程技术研究中心
关键词:
肠出血性大肠杆菌O157∶H7;tox B基因;表达;免疫原性
摘  要:
根据Gen Bank已有tox B基因全长序列,分别设计6对引物扩增tox B基因,克隆到质粒p GEX-4T-1,构建重组菌BL21(plys)(p GEX-4T-1-tox B),IPTG诱导表达重组蛋白质,用Western blot鉴定重组蛋白质免疫原性。tox B基因片段大小分别为1 531 bp、1 590 bp、1 609 bp、1 603 bp、1 525 bp和1 690 bp,重组蛋白质分子量分别为8.1×104、8.3×104、8.4×104、8.3×104、8.0×104和8.7×104。以兔源EHEC O157∶H7抗血清为一抗,Western blot分析结果显示6个重组蛋白质均具有良好的免疫原性。
译  名:
Cloning and expression of toxB gene in segments from enterohemorrhagic Escherichia coli O157 ∶ H7
作  者:
ZHANG Xue-han;LUAN Xiao-ting;HE Kong-wang;NI Yan-xiu;ZHOU Jun-ming;Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences/Key Laboratory of Engineering Research of Veterinary Bio-products,Ministry of Agriculture/National Center for Engineering Research of Veterinary Bio-products;
关键词:
enterohaemorrhagic Escherichia coli O157 ∶ H7;;tox B gene;;expression;;immunogenicity
摘  要:
Based on the tox B sequence on Gen Bank,six primers were designed to amplify the tox B gene.The PCR fragments and p GEX-4T-1 vector were digested with EcoRⅠ and XhoⅠ,ligated with T4 DNA ligase and transformed into the complement BL21(plys) to construct recombinant BL21(plys)(p GEX-4T-1-tox B).After induced by IPTG,recombinant Tox B proteins were expressed.SDS-PAGE analysis revealed that the molecular mass of six recombinant Tox B proteins were8.1×104,8.3×104,8.4×104,8.3×104,8.0×104and 8.7×104,respectively.Western blotting assay using rabbit sera against EHEC O157 ∶ H7 as primary antibody indicated that all six recombinant proteins had good immunogenicities.

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