HBV大分子表面蛋白基因真核重组载体的构建与鉴定  

Cloning and Construction of Human Hepatitis B Virus PreS1/S2/S Gene

作　　者：李志刚[1] 杨林[1] 江元森[1] 顾琳[1] 姚集鲁[1] 

机构地区：[1]中山大学附属第三医院感染科,广州510630

出　　处：《热带医学杂志》2005年第1期19-21,共3页Journal of Tropical Medicine

基　　金：广东省自然科学基金重点项目(No.20013116);广东省自然科学基金(No.04009386);教育部留学回国人员科研启动基金。

摘　　要：目的构建包含人乙型肝炎病毒(HBV)大分子表面蛋白基因的重组载体,以便进一步研究其基因免疫以及对宿主细胞丝裂原活化蛋白激酶(MAPK)p38等信号传导通路的影响。方法设计合成2对寡核苷酸引物,以adr亚型HBV质粒pHBVDNA为模板,采用PCR法分别扩增HBV大分子表面蛋白基因(preS1/S2/S基因)片段;用HindⅢ和BamHI双酶切后,连接到质粒pcDNA3.1(+)相应酶切位点,转化宿主菌DH5α,分别用上述内切酶双酶切及DNA测序鉴定重组质粒。结果酶切鉴定示所切下的片段大小均与预计相符。测序结果与文献报道序列及预计结果一致。结论成功构建了HBV大分子表面蛋白基因的重组载体。Objective To construct recombinant plasmid encoding preS1/S2/S antigen of HBV and to investigate the effect of preS1/S2/S antigen on the mitogen activated protein kinase (MAPK) p38 signaling pathway. Methods The specific primers were designed and synthesized to amplify the DNA fragments encoding preS1/S2/S antigen gene from plasmid pHBV adr by PCR. After being digested with restriction endonuclease, Hind III and BamHI, and purification, the purified preS1/S2/S gene fragments were inserted into pcDNA3.1, and transformed into E.coli DH5α. Then the recombinant plasmid were identified by digesting with restriction endonuclease and DNA sequencing. Results The fragments digested with endonuclease were as large as the predicted results while its DNA sequencing was the same with the reported sequence. Conclusion The recombinant plasmid pcDNA3.1 S1/S2/S was successfully constructed.

关 键 词：乙型肝炎病毒 大分子表面蛋白 重组载体 

分 类 号：R512.62[医药卫生—内科学]