文献类型：期刊文献

中文题名：大肠杆菌腺苷脱氨酶基因的克隆表达

英文题名：Cloning and Expression of E. coli Adenosine Deaminase Gene

作者：栾海涛[1];丁庆豹[1];欧伶[1];魏晓琨[2];许彦梅[2]

机构：[1]华东理工大学生物反应器工程国家重点实验室,上海200237;[2]上海璞诚生物科技有限公司,上海200235

年份：2010

卷号：26

期号：3

起止页码：95

中文期刊名：生物技术通报

外文期刊名：Biotechnology Bulletin

收录：CSTPCD;;北大核心:【北大核心2008】；CSCD:【CSCD_E2011_2012】；

语种：中文

中文关键词：大肠杆菌;腺苷脱氨酶;基因工程

外文关键词：Escherichia call Adenosine deaminase Gene engineering

摘要：将大肠杆菌K12菌株来源的腺苷脱氨酶基因(add)克隆到载体pET-28a中,并转化至大肠杆菌BL21(DE3)中进行表达。通过IPTG诱导,SDS-PAGE检测和酶活性的测定发现,重组菌表达产生大量腺苷脱氨酶,活性达到51.07U/mg蛋白。通过酶性质的研究,腺苷脱氨酶对腺苷最适pH和温度分别为7.5和40℃,且在40℃下维持稳定。
The gene encoding adenosine deaminase（add）from Escherichia coli K12 was cloned into expression vector pET-28a and transformed into the strain E. coli BL21 （DE3）. The protein was highly expressed after the induction with IPTG,then was analyzed with SDS-PAGE and confirmed by the assay of activity. The activity of adenosine deaminase expressed could reached up to 51.07 U/mg protein. The study of characterization showed that adenosine deaminase had optimum pH 7.5 and optimum temperature at 40℃ with thermal stability at 40℃.