枯草芽孢杆菌精氨酸脱羧酶基因speA的表达与蛋白纯化

发布时间：2018-03-12 09:01

  本文选题：枯草芽孢杆菌　切入点：精氨酸脱羧酶　出处：《中国酿造》2017年03期 　论文类型：期刊论文

【摘要】：根据枯草芽孢杆菌(Bacillus subtilis)BJ3-2的精氨酸脱羧酶(ADC)的编码基因spe A序列设计特异性酶切引物,克隆基因speA序列。测序结果显示,基因speA全长为1473 bp,编码490个氨基酸,分子质量为58 ku。基因spe A克隆至原核表达载体,获得重组菌pET28a-spe A/BL21,十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)结果显示,1.0 mmol/L的异丙基β-D-硫代半乳糖苷(IPTG)28℃诱导4 h,上清液和菌体均能表达出ADC蛋白,上清液经纯化、透析、冷冻干燥可获得纯度97%的ADC酶,酶联免疫吸附检测(ELISA)ADC酶活为16780 U/mg。为speA基因的表达、纯化及酶学性质研究奠定了理论基础。
[Abstract]:According to the speA sequence of the arginine decarboxylase of Bacillus subtilis)BJ3-2, the specific primers were designed and the speA sequence of the gene was cloned. The results showed that the length of the gene speA was 1473 BP, encoding 490 amino acids. The molecular weight was 58 ku. the gene spe A was cloned into prokaryotic expression vector. The results of SDS-PAGE showed that the isopropyl 尾 -D- thiogalactoside (IPTG) of 1.0 mmol/L was induced at 28 鈩，

本文编号：1600873