摘要
以大肠杆菌基因组DNA为模板,设计引物扩增得到天冬氨酸酶基因,将其重组于胞内融合表达型T载体中,重组质粒转化表达宿主大肠杆菌BL21(DE3)。SDS-PAGE分析表明,工程菌经IPTG诱导,表达大量表观分子量约75kD的融合蛋白。经试验,工程菌细胞具有较高的天冬氨酸酶活性,融合形式的酶最适温度37℃,最适pH8.5,融合伴侣DsbA的存在对酶活没有影响。
Aspartase gene was amplified and isolated from E.coli genomic DNA by PCR technology,then cloned into the intracellular fusion type T vector,and the recombinant plasmid was transformed into expression host E.coli BL21（DE3）.The IPTG induced engineered strain was analyzed by SDS-PAGE,which showed a fusion protein was expressed with an apparent molecular weight of 75 kD.The engineered cells was approved to have a high aspartase activity,and the recombinant aspartase fusion protein exhibited optimal activity at 37℃ and pH8.5,while the DsbA fusion partner had no affect on the aspartase activity.
出处
《氨基酸和生物资源》
CAS
2012年第1期13-15,20,共4页
Amino Acids & Biotic Resources
