乙型肝炎病毒X基因cDNA的克隆及真核表达载体的构建  

Cloning of human HBx-cDNA and construction of the eukaryotic expression vector

作　　者：尤红娟[1] 裴冬生[2] 孙伟[1] 王慧[1] 

机构地区：[1]徐州医学院微生物与免疫学教研室,江苏徐州221002 [2]徐州医学院生物化学与分子生物学研究中心

出　　处：《徐州医学院学报》2006年第3期191-193,共3页Acta Academiae Medicinae Xuzhou

基　　金：江苏省高校自然科学研究计划(02KJB310010);徐州医学院院课题(04KJ19)基金资助项目

摘　　要：目的为研究HBV X基因(HBx)与原发性肝细胞癌发生的关系,克隆HBx的cDNA并构建HBx真核表达载体。方法利用逆转录聚合酶链反应(RT-PCR)方法从HepG2.2.15细胞中扩增出HBx的cDNA,克隆到质粒PCDNA3.1中。结果RT-PCR扩增出约460 bp片段,经酶切鉴定显示HBx的cDNA真核表达载体(HBx-PCDNA3.1)构建成功,测序结果显示HBx的cDNA与已发表的的序列相同。结论成功地构建了HBx的真核表达载体HBx-PCDNA3.1。Objective To clone the human HBx - cDNA and construct the eukaxyotic expression vector of HBx cDNA for exploring the contribution of HBX gene to the genesis of hepatocellular carcinoma. Methods The total RNA extracted from HepG2.2.15 cells was processed by RT - PCR to produce HBx - cDNA. The product was confirmed in sequence, and then inserted into the et/karyotic expression vector PCDNA3.1. The recombinant plasmid was cloned and designated as HBx - PCDNA3.1. Results A 460 bp cDNA fragment was obtained by RT- PCR. The presence of HBx - cDNA in the vector was confirmed by electrophoresis after double digestion with EcoR Ⅰ and Hind Ⅲ. The sequence of the HBx - cDNA obtained was conistent with the published. Conclusion The task of cloning the HBx - cDNA into the eukaryotic expression vector PCDNA3.1 is accomplished.

关 键 词：乙型肝炎病毒 X基因 克隆 真核表达载体 

分 类 号：R373.21[医药卫生—病原生物学] Q789[医药卫生—基础医学]