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目的构建含乙型肝炎病毒(HBV)表面抗原基因(S区基因)与丙型肝炎病毒(HCV)核心抗原基因(C区基因)的嵌合真核表达载体,观察preS1和preS2基因对HBV表面抗原及HCV核心抗原体液免疫的影响。方法用PCR方法,分别扩增HBVS区基因和HCVC区基因。将S区基因克隆入真核表达载体pcDNA3.1,酶切鉴定后,再将C区基因克隆至S区基因的下游。测序正确后,大量提取质粒并免疫Balb/c小鼠,用ELISA法检测抗HBs和抗HCV抗体。结果成功地扩增出目的基因片段,克隆后酶切鉴定结果正确,序列分析与文献报道相一致。免疫后检测到抗HBs和抗HCV抗体。preS1与preS2基因对构建的融合DNA疫苗的体液免疫应答有一定的抑制作用。抗HBs抗体的产生低于只含S基因的真核表达载体;preS1基因对抗HCV抗体的产生具有抑制作用,而preS2无影响。结论不同长度的HBVS区基因可影响抗HBs和抗HCV抗体的产生。
Objective To construct a chimeric eukaryotic expression vector containing hepatitis B virus surface antigen gene (S region gene) and hepatitis C virus (HCV) core antigen gene (C region gene), and to observe the effect of preS1 and preS2 gene on HBV surface Effect of antigens and HCV core antigen on humoral immunity. Methods The genes of HBVS and HCVC were amplified by PCR. The S region gene was cloned into the eukaryotic expression vector pcDNA3.1. After digestion and identification, the C region gene was cloned downstream of the S region gene. After sequencing, the plasmids were extracted and immunized Balb / c mice, and anti-HBs and anti-HCV antibodies were detected by ELISA. Results The target gene fragment was successfully amplified. After cloning, the result of enzyme digestion was correct, and the sequence analysis was consistent with that reported in the literature. Anti-HBs and anti-HCV antibodies were detected after immunization. PreS1 and preS2 gene fusion DNA vaccine constructed humoral immune response to some extent. The production of anti-HBs antibody was lower than that of eukaryotic expression vector containing only S gene. The preS1 gene had an inhibitory effect on the production of anti-HCV antibody, while preS2 had no effect. Conclusion Different lengths of HBVS gene can affect the production of anti-HBs and anti-HCV antibodies.