摘要
目的构建乙型肝炎病毒X基因原核表达质粒pQE30-HBx,以进一步研究其基因产物的功能及致病机制。方法以肝炎患者血清DNA为模板,用PCR方法扩增乙型肝炎病毒X基因,构建乙型肝炎病毒X基因原核表达质粒pQE30-HBx,经酶切电泳验证重组质粒的正确性,并对X基因进行测序鉴定。结果PCR结果显示,扩增片断大小约465 bp,与预期相同。重组质粒经酶切电泳判断有目的片段插入,方向正确,经测序证实插入片段是乙型肝炎病毒X基因,编码框无误。结论成功构建乙型肝炎病毒X基因重组质粒pQE30-HBx。
Objective To construct the prokaryotic expression plasmid for the X gene of hepatitis B virus （HBx） .Methods The X gene of hepatitis B virus was amplified from serum of hepatitis B patients by PCR method, and inserted into prokaryotic expression vector pQE30;The structure of recombinant pQE30-HBx plasmid was confirmed by restrict endonuclease digestion and sequencing analysis. Results The 465bp DNA fragment of X gene was obtained and inserted exactly in recombinant pQE30-HBx plasmia. Sequencing confirmed that the cloned gene was HBx. Conclusion The recombinant pQE30-HBx plasmia had been constructed successfully.
出处
《哈尔滨医科大学学报》
CAS
北大核心
2007年第3期221-223,共3页
Journal of Harbin Medical University
