当前位置: 首页 > 文章 > 吸水链霉菌TetR基因的克隆与表达 吉林农业大学学报 2004,26 (3) 47-50
Position: Home > Articles > Cloning and Expression of the Gene Encoding Tetracycline Resistance Repressor in Streptomycesis hygroscopius Journal of Jilin Agricultural University 2004,26 (3) 47-50

吸水链霉菌TetR基因的克隆与表达

作  者:
王恒安;秦磊;庞志轩;赵国屏
单  位:
上海交通大学农业与生物学院;中国科学院上海生物工程研究中心
关键词:
吸水链霉菌;四环素抗性阻遏蛋白;蛋白表达
摘  要:
根据吸水链霉菌"应城变种10-22"的一个文库质粒pHZ1392的测序结果设计了1对引物,聚合酶链反应扩增编码TetR的663bp基因,利用T载体和蓝白斑筛选构建了克隆质粒T TetR,经序列测定确认扩增序列正确后克隆至pET24a,构建了表达质粒pET24a TetR,再经序列测定确认未发生移码后转化表达宿主菌E coliDL21(DE3),IPTG诱导后的SDS-PAGE分析显示蛋白表达获得成功,并进一步利用Ni-NTA树脂纯化了TetR蛋白。
译  名:
Cloning and Expression of the Gene Encoding Tetracycline Resistance Repressor in Streptomycesis hygroscopius
作  者:
WANG Heng-an~(1,2), QIN Lei~2, PANG Zhi-xuan~2, ZHAO Guo-ping ~2 (1.Agricultural and Biological College, Shanghai Jiaotong University, Shanghai 201101, China; (2. Shanghai) Research Center of Biotechnology, Chinese Academy of Sciences, Shanghai 200233, China)
关键词:
Streptomyces hygroscopicus; tetracycline resistance repressor; protein expression
摘  要:
According to the annotation results of DNA sequence of pHZ1392, a library cosmid of streptomyces hygroscopicus "yingchengenesis 10-22", a pair of primers were synthesized and used to amplify the gene coding for tetracycline resistance repressor with polymerase chain reaction, the purified DNA fragment was inserted into T vector to construct the cloning plasmid T-TetR. After being sequenced and ascertained correctly, the gene was then inserted into pET24a to construct the expression plasmid pET24a-TetR, the expression host strain E.coli DL21(DE3) was transformed by pET24a-TetR and induced with isopropyl-beta-D-thiogalactopyranoside (IPTG ). The analysis result of SDS-PAGE showed that the TetR was successfully expressed and purified with Ni-NTA resin.

相似文章

计量
文章访问数: 3
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊