摘要
目的：为了获得NDRG3(N-Myc downstream regulated gene3)的cDNA,以成人脑组织为模板。方法：由特异性的引物通过RT—PCR方法扩增人NDRG3的编码基因,大小约为1．1 kb。将此基因插入PMD 18-T载体中,并进行酶切鉴定和序列测定。然后,将测序正确的片段亚克隆人原核表达载体pPROEX-HTb表达载体中,转化DH5α感受态细胞,并在LB培养基中获得高效表达。结果：这表明成功地进行了人NDRG3的克隆和表达,为进一步研究NDRG3的功能奠定了良好的基础。
Objective：To obtain the cDNA of human NDRG3 （N- Myc downstream regulated gene3）.Methods：cl）NA tragment was amplifed by RT - PCR method with specific primers from tissue of human brain. The 1.1 kb fragment was inserted into PMD18 - T vector and identified by di- gestive enzyme and sequencing methods. Then the NDRG3 cDNA was subcloned into prokaryotic expression vector pPROEX- HTb and induced the high expression in DHSα competent cell.Results： These results show that we get the cDNA and the expression of human NDRG3. Thus, it is very helpful to the functional research work of human Ndrg3.
出处
《生物技术》
CAS
CSCD
2006年第5期1-3,共3页
Biotechnology
基金
国家"973"计划项目(2002CB513007)
国家自然科学基金项目资助(30370315
30570676)
教育部长江学者与创新团队发展计划项目(PCSIRT0459)
陕西省自然科学基础研究计划项目(2006C_257)
第四军医大学优秀博士课题资助项目(2004008)
