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.
出处
《徐州医学院学报》
CAS
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
