猪白血病抑制因子(LIF)的原核表达

The Expression of Porcine Leukemia Inhibitory Factor in E.coli. BL21

根据GenBank中猪LIFcDNA基因序列扩增出目的基因片断,并将扩增的片断克隆进pET-31b表达载体的NdeⅠ和XhoⅠ酶切位点之间,经转化使其在大肠杆菌BL21中表达,产生重组猪LIF蛋白。结果表明,所构建的原核表达质粒PET-31b-mpLIF含有编码成熟型猪LIF蛋白的cDNA序列;表达结果显示,诱导样品在相对分子质量约20000的位置上有明显的增强表达条带。

The eDNA fragment encoding mature porcine LIF protein was amplified by pCR according to GenBank. subse- quently cloned into the prokaryotic expression vector PET-31 b at restriction sites Nde I and Xho I, and then transformed into E.coli BL21 for expression.The result show that, the constructed expression plasmid was identified by DNA sequencing and restriction enzyme digestion. The expression of the reconthinant porcine LIF protein was performed exactly in E.coli BL21 and the size was about 20kDa.