稳定表达HBsAg的P815细胞株的建立及鉴定  

Establishment and identification of P815 cell line stably expressing HBsAg

作　　者：周凤娟[1] 戴建新[1] 胡振林[1] 孙树汉[1] 

机构地区：[1]第二军医大学基础医学部医学遗传学教研室,上海200433

出　　处：《第二军医大学学报》2002年第4期384-386,共3页Academic Journal of Second Military Medical University

基　　金：国家"863"计划资助项目(101-06-05-04).

摘　　要：目的：筛选并建立稳定表达亚洲人常见的 adr亚型 HBsAg的 P815细胞株，为乙肝DNA疫苗的效果评价提供体外细胞感染模型。方法：用 PCR方法扩增乙肝病毒的S基因片段后装入PCDNA3载体，并通过脂质体转染 P815细胞，G418筛选。结果：构建了重组质粒pcDNA3－HBsAg（S），转染细胞及其培养上清中均可检测到HBsAg（S）的存在，PCR扩增也证实HBSAg（S）基因已稳定整合于p815细胞的染色体中。结论：获得了稳定表达HBsAg的P815细胞株，为今后在小鼠体内检测乙肝DNA疫苗激发的CTL反应奠定了基础。Objective: To screen for and establish P815 cell line stably expressing HBsAg (adr subtype) which is com-mon in Asian people, providing the cell infection model for evalution of the hepatitis B DNA vaccine effect in virto. Methods:S gene of hepatitis B virus was amplified and cloned into pcDNA3. The recombinant plasmid was transfected P815 by lipo-some and screened by G418. Results: Recombinant plasmid pcDNA3-HBsAg(S) coding hepatitis B (adr subtype) surface pro-tein S was constructed. HBsAg was detected both in the trans fected cell and its culture medium. It was confirmed by PCRthat HBsAg(S) gene was integrated into the chromosome of P8l5 stably. Conclusion: P815 cell line stably expressing HBsAgis obtained, providing bases for detecting the CTL reaction stimulated by hepatitis B DNA vaccine.

关 键 词：乙肝DNA疫苗 体外细胞感染 PCR方法 乙型肝炎表面抗原 ADR亚型 P815细株 

分 类 号：R512.62[医药卫生—内科学]