论文部分内容阅读
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 Lactococcus lactis capable of expressing Salmonella paratyphi A (Pag C). Methods The pag C gene was amplified by PCR, subcloned into p MD18-T vector and sequenced. The pag C gene was digested with restriction endonuclease and inserted into p MG36e and named p MG36e-pag C for transformation into L. lactis MG1363. Recombinant Lactococcus lactis was broken by lysozyme and sonication. The expression of Pag C protein was detected by SDS-PAGE and Western blot. As a result, p MG36e-pag C was identified correctly and Pag C protein expressed by p MG36e-pag C / MG1363 was detected by Western blot. Conclusion The recombinant Lactococcus lactis that can constitutively express Pag C protein was successfully constructed.