利用pET-20b克隆与表达多功能半纤维素酶  

Cloning and Expression of a Trifunctional Hemicellulase Using pET-20b in Escherichia coli

作　　者：彭静静[1] 

机构地区：[1]泰山学院生物与酿酒工程学院,山东泰安271021

出　　处：《湖北农业科学》2014年第17期4205-4207,共3页Hubei Agricultural Sciences

基　　金：泰安市科技发展计划项目(20132094);泰山学院博士科研启动金项目(Y-01-2013001)

摘　　要：将来源于嗜热厌氧乙醇菌(Thermoanaerobacter ethanolicus,JW200)的双活性阿拉伯/木糖苷酶(XarB)构建到pET-20b(+)上,得到质粒pET-20b-xarB。将来源于疏棉状嗜热丝孢菌(Thermomyces lanuginosus DSM 5826)木聚糖酶A(XynA)构建到pET-20b-xarB上,得到表达质粒pET-20b-xarB-xynA。将重组质粒pET-20b-xarB-xynA转入大肠杆菌Escherichia coli JM109(DE3)进行表达。SDS-PAGE结果显示,该重组酶的分子量为108 kDa,与理论值相符。The structure gene from Thermoanaerobacter ethanolicus JW200 encoding arabinosidase-xylosidase XarB gene was amplified and ligated into pET-20b （＋）, resulting in pET-2Ob-xarB. The structure gene from Thermomyces lanuginosus DSM 5826 encoding xylanase（XynA） was amplified and ligated into pET-20b-xarB, resulting in pET-20b-xarB-xynA. The trifunc- tional enzyme （XarB-XynA） was obtained after expression pET-20b-xarB-xynA in Escherichia coli JM109（DE3）. SDS-PAGE analysis showed that the molecular mass of the expressed recombinant XarB-XynA was about 108 kDa, the same as the pre- dicted size.

关 键 词：阿拉伯/木糖苷酶 木聚糖酶 pET-20b(+) 多功能半纤维素酶 克隆 表达 

分 类 号：Q785[生物学—分子生物学]