摘要
目的 希望通过融合表达乙型肝炎病毒前 S1抗原 (pre S1Ag) (1- 42 )及核心抗原 (HBc Ag) (1- 144 )提高预防性疫苗的细胞免疫活性 ,为开发 HBV治疗性疫苗探索一条有效途径。方法 采用 PCR法获得在 HBc Ag(1-144 )基因后连接有 pre S1Ag(1- 42 )基因的融合基因 ,然后将其插入表达载体 p ET- 11d中 ,在大肠杆菌中进行表达。结果 获得了 HBc Ag(1- 144 )与 pre S1Ag(1- 42 )融合蛋白的表达产物 ,SDS- PAGE电泳显示表达产物相对分子质量为 2 0 0 0 0左右 ,约占细菌总蛋白的 2 0 %。经 Western- blot验证 ,表达产物能分别特异地与核心抗体及前 S1抗体结合。经 DNA序列测定 ,pre S1Ag(1- 42 )基因正确地融合在 HBc Ag(1- 144 )基因之后。诱导表达的菌体的裂解液经免疫电镜观察 ,可见到成堆聚集的典型 HBc Ag颗粒。结论 该融合蛋白的表达将为研究其诱导细胞免疫和体液免疫反应的能力 ,以及为今后治疗慢性乙型肝炎的 T细胞疫苗的研制提供有用的资料。
To study the therapeutic T cell vaccine for the treatment of chronic hepatitis B. Methods The genes of HBcAg(1-144) and preS1 Ag(1-42) were amplified and fused by PCR.This fusion gene was inserted in the prokaryotic expression vector pET-11d and expressed in E.coli.Results It was showed by SDS-PAGE that the protein molecular weight of the coexpression product was about 20 000,twenty percent of all bacteria protein.The monoclone antibody against Core and preS1 antigen could react with this fused protein by Western-blot technique respectively.The fused gene was verified by sequencing. Under the immune electron microscopy,this fused protein is a typical particle of HBcAg but in an aggregated form. Conclusions The results may aid for studying T cell immunotherapeutic vaccine to chronic hepatitis B.
出处
《中国医学科学院学报》
CSCD
北大核心
2000年第3期227-231,共5页
Acta Academiae Medicinae Sinicae
