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人铜锌超氧化物歧化酶基因的克隆和乳酸乳球菌中的表达
引用本文:向华,郭顺星.人铜锌超氧化物歧化酶基因的克隆和乳酸乳球菌中的表达[J].生物工程学报,2000,16(1):6-9.
作者姓名:向华  郭顺星
作者单位:[1]中国科学院微生物研究所,北京 [2]中国医学科学院药用植物研究所,北京
基金项目:国家自然科学基金(39800080)和中国博士后科学基金资助。
摘    要:采用RT-PCR技术从人肝总RNA中分离扩增了0.45kb的人铜锌超氧化物歧化酶(Cu/ZnSOD)基因的cDNA序列,首先克隆至大肠杆菌表达质粒pET23b,进行了序列测定和超高表达,将Cu/Zn,SODcDNA亚克隆至乳酸乳球菌表达载体pMG36e,用电穿孔法将重组质粒pMG36esod转化到乳酸乳球菌,获得Cu/Zn SOD的组成型表达,其表达量约占乳酸乳球菌可溶性蛋白的5%以上,活性染色表

关 键 词:超氧化物歧化酶  基因克隆  基因表达  乳酸乳球菌
文章编号:1000-3061(2000)01-0006-04
修稿时间:1999-02-01

Cloning and Expression of Human Cu/Zn Superoxide Dismutase Gene in Lactococcus lactis
XIANG Hua ,WEI Wen-zhong ,TAN Hua-rong GUO Shun-xing.Cloning and Expression of Human Cu/Zn Superoxide Dismutase Gene in Lactococcus lactis[J].Chinese Journal of Biotechnology,2000,16(1):6-9.
Authors:XIANG Hua  WEI Wen-zhong  TAN Hua-rong GUO Shun-xing
Institution:XIANG Hua ,WEI Wen-zhong ,TAN Hua-rong ;(lnstitute of Microbiology, The Chinese Academy of Sciences, Beijing 100080);GUO Shun-xing ;(Institute of Medical Plant, Chinese Academy of Medical Sciences, Beijing 100094)
Abstract:The cDNA encoding human Cu/Zn SOD was amplified by RT-PCR using the total. RNA of human liver as the template, and was cloned into an E. Coli expression vector pET23b. After the DNA sequence was determined, the recom binant plasmid pET23bsod was introduced into E. Coli BL21 (DE3)/pLysS. SDS-PAGE analysis revealed that the recom binant E. Coli expressed the predicted 19 kDa human Cu/Zn SOD, and its amount was over 50% of total proteins. The Cu/Zn SOD eDNA was then subcloned into a lactoeoccal expression vector pMG36e, and resulting pMG36esod was intro duced into L. Lactis MG1363 by electroporation. The human Cu/Zn-SOD was expressed up to 5 % of the soluble proteins,and the enzymatic activity was also observed by SOD activity dying.
Keywords:Cu/Zn-SOD  gene cloning and expression  Lactococcus lactis
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