摘要
采用PCR技术以大肠杆菌JM109基因组DNA为模板扩增得到木糖异构酶基因xylA,连接到载体pET-22b(+),得到重组质粒pET-22b(+)-xylA。将此重组质粒转化到大肠杆菌菌株BL21(DE3)中,重组菌株经IPTG诱导后,通过半胱氨酸-咔唑法测得木糖异构酶活力。每mL发酵液中重组菌株显示出酶活力约为0.84 U。SDS-PAGE电泳结果显示出明显的5×104(相对分子质量)特异性蛋白质条带。
The xylose isomerase gene of E. coli were successfully amplified by PCR reaction and were cloned in vector pET-22b（ ＋ ）. The recombinant vector pET-22b（ ＋ ）-xylA were transformed into E. coli BL21 （DE3）. After induction of IPTG the recombinant xylose isomerase activity was determined by the modified Cysteinearbazole assay. The isomerase activity of broth given by the recombinant stain reached 0.84 U/mL. The specific protein band about 50 kD was showed in the SDS-PAGE gel.
出处
《生物加工过程》
CAS
CSCD
2005年第4期45-48,共4页
Chinese Journal of Bioprocess Engineering
基金
国家"973"项目(编号为2003CB716000)
