水生栖热菌FL-03海藻糖合酶基因克隆及原核表达

Cloning of trehalose synthase gene from Thermus aquaticus FL-03 and prokaryotic expression

海藻糖合酶(Trehalose synthase,TreS)属于α-淀粉酶家族(α-amylase family),是生物体内通过TreS途径生成海藻糖的一种酶类。利用PCR技术,从水生栖热菌FL-03菌株FL-03中扩增了海藻糖合酶基因(TresC01),将其克隆到原核表达载体pET-28a(+),得到重组质粒P-TresC01,转化E.coliBL21(DE3),IPTG诱导表达,重组蛋白经SDS-PAGE分析,得到一条分子量约为110ku的特异条带,并经Western blotting分析鉴定,表明成功构建重组质粒P-TresC01并在大肠杆菌中表达,为进一步的研究奠定基础。

Trehalose synthase pertain α-amylase family,it is a sort of erzymes that producing trehalose by Tres path in biosystem.Trehalose synthase gene（TresC01） was amplified by PCR technique from the trehalose-producing FL-03,that is in the laboratory.TresC01 was cloned into prokaryotic expression vector pET-28a（＋）,obtaining the recombinant plasmid P-TresC01,and expressed in BL21（DE3） under induction of IPTG.The recombination protein is an idio-strap that molecular weight is about 110 ku by the analyses SDS-PAGE.The successed construction recombinant plasmid P-TresC01 by Western blotting identification in order to lay a foundation for further study.