结核杆菌截短型热休克蛋白原核表达的实验研究

An Experimental Study on the Prokaryotic Expression of truncated Heat Shock Protein of Mycobacterium Tuberculosis

探讨结核杆菌的截短型热休克蛋白原核表达技术.以结核杆菌BCG疫苗株为模板,设计PCR引物扩增hsp65基因片段,再以pET32a(+)为载体构建pET-hsp65表达质粒.将pET-hsp65表达质粒转化大肠杆菌BL21,经0.5mM IPTG诱导后,截短型HSP65蛋白与pET32a(+)载体自身的硫氧还蛋白融合表达,融合表达蛋白的分子量约为49.8kDa.

The paper studied the prokaryotic expression technology of the truncated heat shock protein（HSP）of mycobacterium tuberculosis.The BCG vaccine strain was used as a template,and the primers were designed to for amplify hsp65 gene fragment by PCR.Then the expression plasmid of pEThsp65 was constructed by linking hsp65 gene fragment to the vector of pET32 a（＋）.After the pEThsp65 plasmid was transformed into BL21 strain of E.coli by the induction of 0.5 mM IPTG,the truncated HSP65 was successfully fusion-expressed with thioredoxin（TRX）from pET32 a（＋）vector,with a molecular weight of about 49.8 kDa.