| 儿茶酚胺氧位甲基转移酶的克隆表达及纯化 |
| |
| 引用本文： | 李付鸾,谢忠平.儿茶酚胺氧位甲基转移酶的克隆表达及纯化[J].中国生化药物杂志,2007,28(1):17-19. |
| |
| 作者姓名： | 李付鸾 谢忠平 |
| |
| 作者单位： | 中国医学科学院,中国协和医科大学,医学生物学研究所,云南,昆明,650118 |
| |
| 摘 要： | 目的将儿茶酚胺氧位甲基转移酶基因(COMT)克隆到原核表达载体进行可溶性表达,制备纯化COMT蛋白,为深入研究COMT的功能提供材料。方法运用分子生物学方法构建融合表达质粒Pet22b+-COMT。将Pet22b+-COMT转入E.coli BL21(DE3)表达菌株,利用IPTG诱导表达,并用亲和色谱纯化COMT。结果经测序分析成功构建了融合表达质粒Pet22b+-COMT。COMT基因在E.coli BL21(DE3)中表达相对分子质量约为28000的蛋白质,纯化后,COMT蛋白的纯度大于90%。结论COMT基因在原核中得到良好的表达,得到了纯度较高的蛋白质,为酶学活性研究奠定了基础。
|
| 关 键 词： | 儿茶酚胺氧位甲基转移酶 Pet22b+ BL21(DE3) |
| 文章编号： | 1005-1678（2007）01-0017-03 |
| 收稿时间： | 2006-03-02 |
| 修稿时间： | 2006年3月2日 |
Cloning, expression and purification of catechol-O-methyltransferase |
| |
| LI Fu-luan,XIE Zhong-ping.Cloning, expression and purification of catechol-O-methyltransferase[J].Chinese Journal of Biochemical Pharmaceutics,2007,28(1):17-19. |
| |
| Authors: | LI Fu-luan XIE Zhong-ping |
| |
| Institution: | Institute of Medical Biology, Chinese Academy of Medical Science, Peking Union Medical College, Kunming 650118, China |
| |
| Abstract: | Purpose To clon the human soluble catachol-O-Methyltransferase(COMT) into the Pet22b+ vector,expression and purification the COMT protein for researching the function of COMT.MethodsAfter constructing Pet22b+-COMT plasmid,transferred it into E.coli BL21(DE3),then induced by IPTG,and purified by affinity chromatography.ResultsIt is success to build a fusion expresion vector Pet22b+-COMT analyzed by nucleotide sequencing.COMT gene express 28 000 of protein in E.coli BL21(DE3),through puring,the purity of COMT protein can be more than 90%.ConclusionCOMT gene was expressed well in BL21(DE3) E.coli,and highly purified COMT protein are obtained. |
| |
| Keywords: | Pet22b BL21(DE3) |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
