摘要
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。
Aim To construct a set of chimeric plasmids including HBV envelope and HCV core genes and to investigate the effects of preS1 and preS2 on HBV S and HCV C gene expressions. Methods The HBV envelope and HCV core genes were amplified by PCR. The HBV envelope genes was cloned into the eukaryotic expression plasmid pcDNA3.1 and the HCV core gene was cloned into downstream of the HBV envelope gene. The target genes were sequenced and the recombinant plasmids were extracted. After Balb/c mice were immunized with recombinant plasmids and the antibodies against HBsAg and HCV were detected by ELISA. Results The target sequences were amplified and cloned successfully. The sequencing results showed no difference from previous reports. Antibodies against HBsAg and HCV were detected positively in the sera from immunized mice. The anti HCV antibody production was only inhibited by preS1 gene,while the anti HBs antibody production was inhibited by preS1 and preS2 genes. Conclusion It is suggested that different length of HBV envelope genes affect the production of anti HBs and anti HCV antibodies.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2001年第3期234-236,共3页
Chinese Journal of Cellular and Molecular Immunology
