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Thermus thermophilus HB8葡萄糖异构酶在大肠杆菌中表达
引用本文:丁莉,许伟,严明,许琳.Thermus thermophilus HB8葡萄糖异构酶在大肠杆菌中表达[J].生物加工过程,2006,4(2):42-45.
作者姓名:丁莉  许伟  严明  许琳
作者单位:1. 南京工业大学,制药与生命科学学院,南京,210009
2. 南京工业大学,制药与生命科学学院,南京,210009;盐城工学院,化工学院,盐城,224003
基金项目:国家973项目(2003CB716000)
摘    要:为了增加高热稳定性的葡萄糖异构酶的得率,采用PCR技术扩增得到Thermus thermophilusHB8葡萄糖异构酶基因xylA,连接到表达载体pET-22b( )上,获得重组质粒pET-22b( )-xylA。将重组质粒转化到大肠杆菌Rosetta(DE3)中,经IPTG诱导后,通过半胱氨酸-咔唑法测葡萄糖异构酶酶活。重组菌经诱导培养,SDS-PAGE电泳结果显示出明显的分子量约为44 kD特异性蛋白质条带,比酶活约为18.562 U/mg,比野生型菌株提高了2倍。

关 键 词:Thermus  thermophilus  xylA基因  葡萄糖异构酶  克隆  表达
文章编号:1672-3678(2006)02-0042-04
收稿时间:2006-01-20
修稿时间:2006年1月20日

High efficient expression of the glucose isomerase from Thermus thermophilus HB8 in E.coli
DING Li,XU Wei,YAN Ming,XU Lin.High efficient expression of the glucose isomerase from Thermus thermophilus HB8 in E.coli[J].Chinese Journal of Bioprocess Engineering,2006,4(2):42-45.
Authors:DING Li  XU Wei  YAN Ming  XU Lin
Abstract:In order to gain the productivity of glucose isomerase with high thermostability,xylA gene were successfully amplified by PCR reaction from Thermus thermophilus HB8 and were ligated to expression vector pET-22b( ). E.coli Rosetta(DE3) competent cells were transformed with the recombinant plasmid pET-22b( )-xylA.The glucose isomerase activity of recombinant strain was determined after IPTG induction by modified Cysteine-carbazole assay.The special protein band about 44 kD was shown in the SDS-PAGE gel.The glucose isomerase activity of recombinant strain was 18.562 U/mg under optimal conditions and 3 times as much as that of wild-type strain.
Keywords:Thermus thermophilus xylA gene  glucose isomerase  clone  expression
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