水貂IFN-α基因的克隆与原核表达

Cloning and prokaryotic expression of ferret IFN-α genes

从水貂基因组中通过PCR的方法克隆得到干扰素α基因(IFN-α),将IFN-β成熟肽插入原核表达载体pET32a,在E.coliBL21(DE3)工程菌中表达重组蛋白。结果表明IFN-β片段长564 bp,与已报道的水貂干扰素-α基因相比同源性98%。IFN-α成熟肽可编码166个氨基酸。重组质粒经ITPG诱导6 h后蛋白表达产物较高,表达量占菌体表达量的50%,分子量约为36 ku,与预期结果相符。

Interferon-beta gene（IFN-α） of ferret was cloned from ferret genome by PCR.IFN-β mature peptide was inserted into the expression vector pET32a,then transformed into BL21 E.coli strain.The results indicated that the IFN-β fragment was 564bp and the sequencing result showed that there is 98% homology among the documented sequences and sequence reported here.SDS-PAGE confirmed protein expressed highly about 50% of the total host bacterium when the recombinant was induced with IPTG for 6 hours.And the molecular weight of exogenous protein was about 36ku as expected.