| Cloning and Expression of Apolipoprotein E3 and Its Variant apoE2 and apoE4 |
| |
| 作者姓名： | 宗义强 刘志国 毕昊 姚研怡 过健俐 屈伸 |
| |
| 作者单位： | [1]Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Tongji Medical College, Huazhong University of Science and Technology , Wuhan 430030, China [2]Department of Biotechnology and Chemical Engineering, Wuhan Polytechnic University, Wuhan 430023, China |
| |
| 摘 要： | In order to obtain three isoforms of apolipoprotein E （apoE）, the eDNA encoding apoE3 was obtained by RT-PCR from normal human liver tissue, Site-directed mutagenesis was used to obtain the cDNAs encoding apoE2 and apoE4 isoforms. The 3 cDNAs were subcloned into vector pGEM-3Z and verified by DNA sequencing. The expression recombinant which can express the target protein as a （His） 6-tagged fusion was constructed by subcloning apoE eDNA into vector pTT-PL, The purified proteins were gained by Ni-NTA column, The SDS-PAGE results revealed the 6 His fusion proteins （apoE2, apoE3 and apoE4） were correctly expressed and purified successfully.
|
| 关 键 词： | 无性繁殖 基因表达 载脂蛋白E3 apoE2 apoE4 变异体 |
| 收稿时间： | 19 September 2005 |
Cloning and expression of apolipoprotein E3 and its variant apoE2 and apoE4 |
| |
| Zong Yiqiang,Liu Zhiguo,Bi Hao,Yao Yanyi,Guo Jianli,Qu Shen.Cloning and Expression of Apolipoprotein E3 and Its Variant apoE2 and apoE4[J].Journal of Zuazhong University of Science and Technology: Medical Edition,2006,26(1):1-3. |
| |
| Authors: | Zong Yiqiang Liu Zhiguo Bi Hao Yao Yanyi Guo Jianli Qu Shen |
| |
| Institution: | 1. Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
2. Department of Biotechnology and Chemical Engineering, Wuhan Polytechnic University, Wuhan 430023, China |
| |
| Abstract: | Summary In order to obtain three isoforms of apolipoprotein E (apoE), the cDNA encoding apoE3 was obtained by RT-PCR from normal human
liver tissue. Site-directed mutagenesis was used to obtain the cDNAs encoding apoE2 and apoE4 isoforms. The 3 cDNAs were subcloned
into vector pGEM-3Z and verified by DNA sequencing. The expression recombinant which can express the target protein as a (His)
6-tagged fusion was constructed by subcloning apoE cDNA into vector pT7-PL. The purified proteins were gained by Ni-NTA column.
The SDS-PAGE results revealed the 6 His fusion proteins (apoE2, apoE3 and apoE4) were correctly expressed and purified successfully.
Zong Yiqiang, male, born in 1968, Technician
This project was supported by a grant from the National Natural Sciences Foundation of China (No. 30100134). |
| |
| Keywords: | apoE gene cloning site-directed mutation gene expression |
| 本文献已被 维普 万方数据 SpringerLink 等数据库收录！ |
|
