重组奶牛干扰素-α基因的原核表达

Prokaryotic Expression of Cow Interferon-α Fusion Gene

从健康奶牛的血液中分离白细胞,用刺激物诱导白细胞产生干扰素,采用RT-PCR扩增干扰素-α基因,构建原核表达载体pQE30-IFNα,用IPTG诱导表达。得到重组的奶牛成熟干扰素-α基因全长498bp,构建的原核表达载体诱导6h后表达量较高,表达产物的分子量约为20kD,从而获得了表达奶牛干扰素-α的基因工程菌株。

Use the principle of gene engineering, the recombinant plasmid with cow interferon-α fusion gene was constructed, and the interferon-α gene was expressed in Escherichia coli. White ceils separated from vein blood of healthy cow were stimulated with NDV-F and ConA to produce interferon. Through the RT-PCR, the interferon-α gene was cloned, and its length was known as 498bp by sequence analysis. Then it was linked to plasmid pQE30 which was transfected into E.coli BL21 later, and then the BL21 was induced by IPTG. The exogenous protein expressed highly when the recombinant was induced for 6 hours. And the molecular weight of exogenous protein was about 20kDa.