摘要
目的构建结核分枝杆菌LprG基因的融合表达载体,在大肠杆菌中进行表达,为研究其在结核病诊断中的作用奠定基础。方法采用聚合酶链反应(PCR)从结核分枝杆菌H37Rv基因组中扩增出LprG基因,克隆到pEASY T1中,序列测定正确后,再亚克隆到融合表达载体PET28a(+)中,转化入大肠杆菌BL21(DE3),经IPTG诱导表达。结果获得结核分枝杆菌LprG蛋白,凝胶薄层扫描分析表达量在38%左右,主要以可溶性形式存在。结论获得了重组结核分枝杆菌LprG蛋白,为以后研究奠定了基础。
To construct the prokaryotic expression vector for lprG gene from Mycobacterium tuberculosis and to express LprG protein in E. coli,the lprG gene extracted from the genomic DNA of M. tuberculosis H37Rv strain was amplified by PCR and cloned to vector pEASY T1. After sequencing,it was subcloned to fusion expression vector pET28a（＋）and then transfected to E coli BL21 （DE3）. It was found that the LprG protein was successfully expressed after induction with IPTG. As demonstrated by thin layer chromatography scanner, the amount of the protein expression approached up to 38% and mainly existed in soluble form. These results would provide the basis for the further studies of LprG protein.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2007年第10期1013-1015,共3页
Chinese Journal of Zoonoses
