以减毒鼠伤寒沙门氏菌为载体的重组HBsAg口服疫苗构建与鉴定

Construction and Identify of Recombinant Attenuated Salmonella typhimurium Orally HBsAg Vaccine

采用大量培养含有pcDNA3s的大肠杆菌,提取pcDNA3s质粒,利用电转化方法将pcD-NA3s质粒转化到感受态减毒鼠伤寒沙门氏菌,SDS-PAPG检测到930 bp大小的条带,基因序列分析到重组菌含有乙肝表面抗原(HBsAg)基因序列;观察重组菌体外传代培养的稳定性,该重组菌株在体外能稳定地繁殖、生长和传代。试验结果表明,携带HBsAg的重组减毒鼠伤寒沙门氏菌疫苗X8786/pcDNA3s已成功构建,为研究和应用人和动物治疗用乙型肝炎病毒口服基因工程活疫苗奠定了基础。

To construct recombinant attenuated salmonella typhimurium vaccine strain X8786/ pcDNA3s expressing HBsAg, plasmid pcDNA3s from recombinant coli was distilled, electricity transformation method was used in plasmid pcDNA3s transformating to recepted state salmonella typhimurium. SDS-PAPG examined 930 bp strip belt, gene sequence analyzed recombinant bacterium have Hepatitis B Surface Antigen （HBsAg） gene sequence. The stability of recombinant bacterial cultured in vivo was observed. It showed that recombinant bacterium could stably reproduction,growth and hand down from generation to generation in vivo. The results confirmed that recombinant attenuated salmonella typhimurium vaccine strain X8786/pcDNA3s carryng HBsAg was constructed succeed. It established foundation of research and development in therapying humen and animal HBV orally gene engineering live vaccine.