乙型肝炎病毒多表位抗原基因的设计、合成及表达

目的　设计合成乙型肝炎病毒多表位抗原基因并在大肠杆菌中表达 ,以期获得兼具预防和治疗作用的新型乙肝疫苗。方法　设计并合成一条乙型肝炎病毒 (HBV)多表位抗原基因 (P T) ,该基因包括HBV前S2区的核苷酸序列 ,和分别来自乙型肝炎表面抗原、乙型肝炎核心抗原、乙型肝炎DNA聚合酶的其它 4个连续的HLA限制性抗原表位的核苷酸序列。基因合成后克隆入载体质粒pWR4 5 0 1,转化大肠杆菌BL2 1,IPTG诱导P T基因与载体质粒的 β 半乳糖酐酶基因融合表达。结果　成功合成并拼接出乙型肝炎多表位抗原基因P T ,阳性重组子PWR HBV P T构建成功 ,并在大肠杆菌中融合表达出相对分子质量 (Mr)约 75× 10 3的碱性蛋白质。结论　初步设计成功HBV多表位抗原基因 ,在大肠杆菌中可表达出具有良好抗原性的重组融合蛋白 ,可能是较理想的HBV预防、治疗性疫苗候选物。

Design, synthesis and expression of the multi-epitope gene of hepatitis B virus

Objective To study the design, synthesis and expression of the multi-epitope gene of hepatitis B virus and to obtain a new therapeutic and preventive vaccine of hepatitis B infection. Methods To designed and synthesize the multi-epitope gene of hepatitis B virus containing has five epitope genes that comes from pre-S2 region, surface antigen, core antigen and DNA polymerase of HBV virus. The multi-epitope gene was cloned into expression vector pWR450-1 and expressed in Escherichia coli BL21. Results Recombinant plasmid PWR/HBV-P/T was successfully constructed and the protein of multi-epitope gene of hepatitis B virus was expressed in Escherichia coli, with good antigenicity. Conclusion The designed multi-epitope therapeutic gene of hepatitis B virus was proved to be successful and the expressed protein may be a potential candidate of therapeutic and preventive vaccine.