摘要
目的克隆结核分枝杆菌CFP10基因,在大肠杆菌中进行表达,为进一步研究其在结核病诊断中的价值奠定基础。方法以结核分枝杆菌H37Rv基因组DNA为模板,通过PCR方法对CFP10的基因进行扩增,以PET-30a(+)为载体构建重组质粒,将重组质粒先转化入大肠杆菌DH5α中,抽提质粒,进行双酶切鉴定,再转化到表达宿主菌BL21(DE3)中,以IPTG诱导表达,聚丙烯酰胺凝胶电泳(SDS-PAGE)分析蛋白表达形式。结果构建了具有正确基因序列的CFP10重组表达质粒,重组蛋白在大肠杆菌BL21(DE3)中以包涵体形式表达。结论目的基因克隆入宿主菌中并表达成功,为深入研究该蛋白的生物学、免疫学活性奠定了基础。
To clone and express the gene coding for Mycobacterium tuberculosis antigen CFP10, and to establish a basis [or diagnosis of tuberculosis, the gene coding CFP10 was amplified by polymerase chain reaction （PCR）, and then the gene was inserted to vector pET-30a（＋）to construct the recombinant plasmid. The recombinant plasmid was transformed into expressive vector E. coli BL21（DE3）, and was induced with IPTG. The presence of recombinant protein in the expression vector was analyzed by SDS-PAGE. The recombinant CFP10 protein was expressed in inclusion body in E. coli BL21 （DE3）, and the target gene had been cloned into host bacterium. These results would establish a basis for diagnosis of tuberculosis.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2007年第7期703-705,共3页
Chinese Journal of Zoonoses
