Thermus thermophilus HB8葡萄糖异构酶在大肠杆菌中表达  

High efficient expression of the glucose isomerase from Thermus thermophilus HB8 in E.coli

作　　者：丁莉[1] 许伟[1] 严明[1] 许琳[1] 

机构地区：[1]南京工业大学制药与生命科学学院,南京210009

出　　处：《生物加工过程》2006年第2期42-45,共4页Chinese Journal of Bioprocess Engineering

基　　金：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。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 IPTO 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.

关 键 词：THERMUS THERMOPHILUS xylA基因 葡萄糖异构酶 克隆 表达 

分 类 号：Q786[生物学—分子生物学]