鸡白痢沙门氏菌外膜蛋白C基因(OmpC)的克隆与表达

Cloning and expression of the gene encoding the outer membrane proteinC (OmpC) of Salmonella pullorum

以鸡白痢沙门氏菌基因组DNA为模板,采用PCR技术扩增得到1026bp的OmpC基因片段。将扩增的OmpC基因克隆至大肠杆菌-乳酸菌穿梭表达载体pW425et中,构建原核表达重组质粒pW425et-OmpC。将pW425et-OmpC转化至thyA基因缺陷型大肠杆菌感受态E.coliX13中。SDS-PAGE分析可见1条约36ku的融合蛋白。Western blot分析表明,该重组蛋白具有反应原性,本研究结果为pW425et-OmpC在乳酸菌中的表达提供了实验依据。

To clone and express the gene encoded the outer membrane protein C of Salmonella pullorum. The 1 026 bp OmpC gene of S.pullorum was amplified by PCR from the genomic DNA, cloned into pW425et vector and transformed into competent E.coli X13 cells. The recombinant protein was induced to express with IPTG. SDS-PAGE showed a 36 ku protein was expressed and western blot confirmed the antigenicity against positive sera. The recombinant protein could be useful resource for development of genetic engineering Lactobacillus oral vaccine.