| 秀丽隐杆线虫胆固醇7,8-脱氢酶在大肠杆菌中的表达 |
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| 引用本文： | 格日勒,申雁冰,任佳佳,汤睿,王敏.秀丽隐杆线虫胆固醇7,8-脱氢酶在大肠杆菌中的表达[J].天津科技大学学报,2014(5):24-28. |
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| 作者姓名： | 格日勒 申雁冰 任佳佳 汤睿 王敏 |
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| 作者单位： | 工业发酵微生物教育部重点实验室，天津市工业微生物重点实验室，天津科技大学生物工程学院，天津 300457 |
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| 基金项目： | 国家高技术研究发展计划“863计划”资助项目(2011AA02A211)；国家自然科学基金资助项目 |
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| 摘 要： | 胆固醇7,8–脱氢酶是参与胆固醇代谢的还原酶,可以将胆固醇转化为7–脱氢胆固醇(合成维生素D3的重要中间体).合成秀丽隐杆线虫的胆固醇7,8-脱氢酶基因daf-36,与具有T7强启动子的载体pET32a(+)连接后,成功构建pET32a-daf-36表达质粒,转入大肠杆菌表达系统E.coli BL21(DE3)和E.coli Rosetta(DE3),经过IPTG诱导培养,蛋白质电泳图谱及质谱结果表明此酶得到了高效表达.
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| 关 键 词： | 胆固醇7 8-脱氢酶基因daf-36 基因克隆 大肠杆菌 蛋白质表达 |
Expresion of cholesterol 7,8-dehydrogenase of Caenorhabditis elegans in E. coli |
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| GE Rile,SHEN Yanbin,REN Jiajia,TANG Rui,WANG Min.Expresion of cholesterol 7,8-dehydrogenase of Caenorhabditis elegans in E. coli[J].Journal of Tianjin University of Science & Technology,2014(5):24-28. |
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| Authors: | GE Rile SHEN Yanbin REN Jiajia TANG Rui WANG Min |
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| Institution: | GE Rile;SHEN Yanbin;REN Jiajia;TANG Rui;WANG Min;Key Laboratory of Industrial Fermentation Microbiology,Ministry of Education,Tianjin Key Laboratory of Industrial Microbiology,College of Biotechnology,Tianjin University of Science & Technology; |
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| Abstract: | Cholesterol 7,8-dehydrogenase DAF-36 is a key enzyme involved in cholesterol metabolism. In this study,daf-36 gene from Caenorhabditis elegans was synthesized. The subcloned daf-36 pET-32a（＋）was to construct the expression plasmid pET32a-daf-36. The recombined plasmid was then transformed into E.coli BL21（DE3）and E. coli Rosetta（DE3）. The results of SDS-PAGE and MS showed that the enzyme could be efficiently expressed in the recombined bacteria after being induced by IPTG. |
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| Keywords: | cholesterol 7 8-dehydrogenase gene daf-36 gene cloning E coli protein expression |
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