摘要
目的:构建HBVX基因与pCI-neo相融合的高效真核表达载体。方法:设计并合成HBVX基因的引物,以HBVDNA阳性血清PCR扩增得到HBVX基因全序列,将X基因连接到真核表达载体pCI-neo上后,酶切图谱分析、PCR检测和扩增产物序列分析等,鉴定所构建的真核表达载体。结果:以重组载体为模板扩增得到的片段大小与已知HBVX基因大小相同,酶切也得到目的基因片段,测序结果也显示与已知X基因序列相同。结论:成功构建了HBVX基因的真核表达载体,为进一步研究HBVX基因及其产物的功能奠定了基础。
Objective:To construct a highly effective eukaryotic expression vector pCI-neo with HBV X gene(pCI-neo-X).Methods:HBV DNA was extracted from the HBV DNA positive serum.HBV X gene was amplified by PCR,and identified by gelose electrophoresis analysis.The PCR products and the eukaryotic expression vectors pCI-neo were digested by restriction endonucleases,then the digested X gene was inserted into the digested eukaryotic expression vector.After screening,the positive recombinants were picked out.Results:PCR analysis showed that the HBV X gene was successfully inserted into pCI-neo vector.Conclusion:The construction of the eukaryotic expression vector pCI-neo will lay solid foundations for further study of HBVX gene.
关键词
乙型肝炎病毒
X基因
真核表达载体
Hepatitis B virus
X gene
Eukaryotic expression vector
