甲型副伤寒沙门菌PagC蛋白在乳酸乳球菌中的表达研究

Expression of Pag C protein of Salmonella paratyphi A in Lactococcal Lactis

目的构建能够表达甲型副伤寒沙门菌外膜蛋白(Pag C)的乳酸乳球菌。方法 PCR扩增pag C基因,亚克隆至p MD18-T载体并测序,从亚克隆载体中酶切回收pag C基因,插入p MG36e并命名为p MG36e-pag C,转化乳酸乳球菌MG1363。溶菌酶加超声的方法破碎重组乳酸乳球菌,SDS-PAGE以及Western blot检测Pag C蛋白的表达。结果 p MG36e-pag C鉴定正确,Western blot检测到p MG36e-pag C/MG1363表达的Pag C蛋白。结论成功构建能组成型表达Pag C蛋白的乳酸乳球菌。

Objective To construct a recombinant Lactococcal Lactis which could express the outer membrane protein（ Pag C） of Salmonella paratyphi A. Methods The pag C gene was amplified by PCR and subcloned into p MD18-T vector for sequencing. The above fragment was inserted to p MG36 e. The new plasmid was named p MG36e-pag C and transferred to Lactococcal Lactis MG1363. The recombinant p MG36e-pag C / MG1363 were broken by lysozyme digestion and ultrasonication. SDS-PAGE and Western blot were used to detect the expressed protein. Results The p MG36e-pag C was comfirmed by the identification. The expression of Pag C protein was detected in the p MG36e-pag C / MG1363 by Western blot. Conclusion The recombinant bacteria p MG36e-pag C / MG1363 was constructed successfully,which could express Pag C protein.