摘要
利用乳糖诱导重组大肠杆菌BL21(DE3)中Pfu DNA聚合酶基因的表达。对诱导菌株起始生长量、乳糖浓度和诱导持续时间进行了优化,尝试使用JK110弱阳离子交换柱和Sephadex G-75凝胶层析柱来分离纯化Pfu DNA聚合酶,并用PCR扩增实验检测酶活性。结果表明乳糖可以有效诱导Pfu酶的表达,JK110离子交换柱有较好的纯化效果,从500 mL培养液中可以得到3×105U的酶活,比活达22,200 U/mg,结果好于IPTG诱导表达。
The expression of Pfu DNA polymerase in recombinant E. coli BL21 （DE3） induced by lactose was studied in this paper. We optimized the fermentation conditions,including initial biomass of induction,concentration of lactose and lasting time of induction. We also tried to purify Pfu DNA polymerase by JK110 weak acid cationic exchange column and Sephadex G-75 gel chromatography. The activity of purified Pfu DNA polymerase was assayed by PCR amplification. The result indicated that lactose could induce the expression of Pfu DNA polymerase effectively,and JKl10 worked well in the purification. A total of 3 10^5U of polymerase was obtained from 500 ml culture,and its specific activity was 22,200 U/mg. This result was better than that of IPTG.
出处
《科技通报》
2008年第1期23-28,共6页
Bulletin of Science and Technology
