| 人神经营养素-4基因的克隆及表达 |
| |
| 引用本文： | 崔羽,常洪起,路明霞,李景鹏,周文婷,阳燕,王立达.人神经营养素-4基因的克隆及表达[J].哈尔滨医科大学学报,2006,40(5):367-370. |
| |
| 作者姓名： | 崔羽 常洪起 路明霞 李景鹏 周文婷 阳燕 王立达 |
| |
| 作者单位： | 1. 东北农业大学生命科学学院,黑龙江,哈尔滨,150030
2. 哈尔滨接触科技有限公司,黑龙江,哈尔滨,150090 |
| |
| 摘 要： | 目的 通过基因工程的方法扩增人神经营养素-4(hNT-4)cDNA,在大肠杆菌中表达hNT-4,为研究hNT-4的生物学作用提供材料.方法 以人胎脑RNA为模板,采用RT-PCR技术扩增hNT-4 cDNA,将其插入的质粒pGEM-T中.将经过序列分析确定的hNT-4 cDNA插入原核表达载体pGEX-6p-1中,构建重组表达载体pGEX-6p-1-NT-4,用SDS-PAGE法测定融合蛋白在大肠杆菌中的表达.结果 经序列测定分析,克隆的hNT-4 cDNA核苷酸序列与已发表序列(GenBank,M86528)完全一致,并获得了高效表达hNT-4的重组菌株.结论 体外成功扩增hNT-4 cDNA,并在原核细胞中高效表达了hNT-4.
|
| 关 键 词： | 人神经营养素-4 基因克隆 原核表达 |
| 文章编号： | 1000-1905（2006）05-0367-04 |
| 收稿时间： | 2005-10-31 |
| 修稿时间： | 2005年10月31 |
Cloning and expression of the human neurotrophin-4 |
| |
| CUI Yu, CHANG Hong-qi, LU Ming-xia, et al.Cloning and expression of the human neurotrophin-4[J].Journal of Harbin Medical University,2006,40(5):367-370. |
| |
| Authors: | CUI Yu CHANG Hong-qi LU Ming-xia |
| |
| Institution: | Life Sciences College, Northeast Agriculture University, Harbin 150030, China |
| |
| Abstract: | Objective Clone human neurotrophin-4 cDNA through the biotechnical method,express its protein in E.coli,and provide material for researching the biological activity of hNT-4.Methods With the RNA of human fetal brain as template,hNT-4 cDNA was amplified by RT-PCR and recombinated into phage pGEM-T.After being sequenced,the cloned hNT-4 cDNA was recombinated into expressed vector pGEX-6p-1,and the fusion protein was expressed in E.coli and identified by SDS-polyacarylamide gel electrophoresis.Results The sequence of the cloned hNT-4 cDNA was completely the same as the reported one(GenBank,M86528),and the bacterium line which could express the fusion protein efficiently was screened out.Conclusion The hNT-4 cDNA was amplified and hNT-4 was expressed in vitro successfully. |
| |
| Keywords: | human neurotrophin-4/5(NT-4/5) gene cloning prokaryotic expression |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
