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| 胆固醇氧化酶基因的克隆及其在大肠杆菌中的表达 | |
| 引用本文： | 银国利,张锐,郭三堆,陈毓荃,牛天贵.胆固醇氧化酶基因的克隆及其在大肠杆菌中的表达[J].西北农业学报,2003,12(3):12-16,35. |
| 作者姓名： | 银国利 张锐 郭三堆 陈毓荃 牛天贵 |
| 作者单位： | 1. 中国农业科学院生物技术研究所,北京,100081;西北农林科技大学,陕西,杨凌,712100 2. 中国农业科学院生物技术研究所,北京,100081 3. 西北农林科技大学,陕西,杨凌,712100 4. 中国农业大学,北京,100094 |
| 基金项目： | "863"计划项目资助(2001AA222101). |
| 摘 要： | 采用PCR方法从来源于蒙古野驴的马红球菌(Rhodococcus equi)4-2G2菌株中分离出编码胆固醇氧化酶的新基因(choEW),其与已知的胆固醇氧化酶基因choE和choB同源性均达99％;按其推断的氨基酸序列与choE和choB相比在485位缺少一个氨基酸,分析认为choEW与choB和choE属于同一基因家族。将choEW插入到原核表达载体pET-His中,构建出重组质粒pETW,并转化Escherichia coli BL21(DE3)plysS获得工程菌。经IPTG诱导后表达出分子量约为56 kD的蛋白质,与核苷酸序列推断的蛋白大小相符。目的蛋白表达量约占细胞总蛋白的11.9％。 |
| 关 键 词： | 胆固醇氧化酶 基因克隆 choEW 基因表达 |
| 文章编号： | 1004-1389(2003)03-0012-05 |
| 收稿时间： | 2003/5/26 0:00:00 |
| 修稿时间： | 2003/6/26 0:00:00 |
Cloning of Cholesterol Oxidase Gene from Rhodococcus Equi and Its Expression in Escherichia Coli |
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| YIN Guo-li,ZHANG Rui,GUO San-dui,CHEN Yu-quan and NIU Tian-gui.Cloning of Cholesterol Oxidase Gene from Rhodococcus Equi and Its Expression in Escherichia Coli[J].Acta Agriculturae Boreali-occidentalis Sinica,2003,12(3):12-16,35. | |
| Authors: | YIN Guo-li ZHANG Rui GUO San-dui CHEN Yu-quan and NIU Tian-gui |
| Institution: | Biotechnology Research Institute, CAAS, Beijing 100081, China;Northwest Scie-Tech University of Agriculture and Forestry, Yangling Shaanxi 712100, China;Biotechnology Research Institute, CAAS, Beijing 100081, China;Biotechnology Research Institute, CAAS, Beijing 100081, China;Northwest Scie-Tech University of Agriculture and Forestry, Yangling Shaanxi 712100, China;China Agricultral University, Beijing 100094, China |
| Abstract: | In this report,a new gene,choEW,encoding a cholesterol oxidase was cloned from Rhodococcus equi using PCR methods,which showed 99% homology to the previously reported cholesterol oxidase genes choE and choB;The putative ORF(opening reading frame) lacks an amino acid residue at 485 site.Analyses showed that choEW belongs to the same gene family as choE&choB do.The choEW was inserted into prokaryotic expression vector pET-His,and the resulting recombinant plasmid pETW was used to transform E.coli BL21(DE3)plysS.A 56 kD protein product was detected using SDS-PAGE after IPTG induction,the size of which consisted with the molecular weight of the putative amino acid.The expressed product accounted for about 11.9% of the total cell protein. |
| Keywords: | Cholesterol oxidase Gene cloning choEW Gene expression |
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