人NDRG4-BcDNA克隆和表达  

Cloning and expression of human NDRG4-B cDNA

作　　者：张健[1] 王立峰[1] 苏金[1] 刘新平[1] 张万会[2] 药立波[1] 

机构地区：[1]第四军医大学基础部生物化学与分子生物学教研室,陕西西安710033 [2]第四军医大学基础部生理学教研室,陕西西安710033

出　　处：《第四军医大学学报》2003年第13期1218-1220,共3页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金 (30 0 70 773 ;30 1 71 0 4 4 )

摘　　要：目的 :获得NDRG4 B(N mycdownstream regulatedgene 4 B)的编码基因 ,表达NDRG4 B蛋白 .方法 :以成人脑cDNA文库为模板 ,通过PCR方法扩增NDRG4 B编码基因 ,克隆至pMD18 T载体并测序 ,再亚克隆至表达载体pRSET A ,转化大肠杆菌 ,IPTG诱导表达 6His NDRG4 B融合蛋白 .结果 :从成人脑cDNA文库中扩增出编码NDRG4 B的cDNA ,序列测定证实有两个沉默突变 (从ATG开始 ,第 76 5bp处 :G→A ;96 6bp处 :A→G) ;成功构建了 pRSET A融合表达载体NDRG4 B pRSET A ;表达了 6His NDRG4 B融合蛋白 .结论 :克隆与表达了人NDRG4 B .AIM: To clone the cDNA of human NDRG4 B and to express the corresponding protein. METHODS: PCR was used to amplify the NDRG4 B coding region from human brain cDNA library. The PCR product was cloned into pMD18 T plasmid, sequenced and then subcloned into vector pRSET A. The NDRG4 B protein was expressed in E. coli of BL21 as a fusion protein with six histidine residues induced by IPTG. RESULTS: NDRG4 B coding region was cloned into pRSET A and the sequence was confirmed to harbor two silent mutations (the 765th bp and 966th bp after the starting codon: G→A and A→G respectively). Fusion expressing vector of NDRG4 B pRSET A was successfully constructed and 6His NDRG4 B fusion protein was correctly expressed. CONCLUSION: Human NDRG4 B was successfully cloned and expressed.

关 键 词：NDRG4-B N-MYC 聚合酶链反应 基因克隆 融合表达 基因表达 

分 类 号：Q78[生物学—分子生物学]