鼠疫耶尔森氏菌LcrV基因在大肠杆菌中的高效表达

High-level expression of Yersinia pestis LcrV gene in Escherichia coli

将测序后的鼠疫耶尔森氏菌 (Yersiniapestis)LcrV基因重组质粒pGEM T ypV酶切 ,克隆于原核表达载体pBV2 2 0 ,构建成pBV ypV表达质粒 ,转化大肠杆菌DH5α ,进行PCR及酶切鉴定 ,筛选阳性克隆 ,进行温控诱导表达 ,SDS PAGE检测表达产物 ,在相对分子质量 3 80 0 0处有一表达条带 ,经薄层扫描分析目的蛋白条带占全菌蛋白的3 8.4 %以上 ,主要以可溶形式存在。

The recombinant expression plasmid pBV/ypV was constructed by inserting the DNA fragment of Yersinia pestis LcrV gene into pBV220 and transformed into E.coli DH5α cell. The positive clones were screened out by PCR and enzymes digesting, and then through temperature regulated inducement expression, the expressed product was detected by SDS PAGE, and an expression band about 38000 was found. The aim protein representing 38.4% of total cell protein and the most in soluble form.