论文部分内容阅读
目的 :针对我国乙型肝炎 (乙肝 )病毒流行的血清型 ,采用美国药品及食物鉴定委员会承认的应用于疫苗临床使用的载体 p VAX1,构建乙肝病毒 adw血清型核酸疫苗 pre S2 S的真核表达质粒 ,对其在真核细胞中的表达进行初步研究。方法 :采用 PCR法扩增 pre S2 S片段 ,插入 p VAX1载体中 ,测定 DNA序列后 ,转染 SP2 / 0细胞。抽提转染后 SP2 / 0细胞的总 RNA ,再用 RT- PCR法扩增其中的 pre S2 - S片段 ,以检测其表达的基础。结果 :测序证实了该片段为乙肝病毒 adw型 pre S2 - S基因。PCR扩增法检测出的转染细胞中存在 pre S2 S基因。结论 :获得了 adw血清型乙肝病毒 pre S2 S的真核表达质粒 ,提示其能够在真核细胞中表达目的基因蛋白
OBJECTIVE: To detect the prevalence of hepatitis B virus (HBV) serotypes in China by using the vector p VAX1 approved by the US Medicines and Food Evaluation Committee for the clinical use of the vaccine and to construct the eukaryotic expression vector of preS2 Plasmids, their expression in eukaryotic cells for a preliminary study. Methods: Pre S2 S fragment was amplified by PCR and inserted into p VAX1 vector. The DNA sequence was determined and transfected into SP2 / 0 cells. The total RNA of SP2 / 0 cells was extracted and the pre S2 - S fragment was amplified by RT - PCR to detect the expression of SP2 / 0 cells. Results: Sequencing confirmed that the fragment was HBV preW - adw gene. Pre S2 S gene is present in transfected cells detected by PCR amplification. Conclusion: The eukaryotic expression plasmid of adw serogroup HBV pre S2 S was obtained, suggesting that it can express the target gene protein in eukaryotic cells