甲型肝炎病毒编码区cDNA在减毒伤寒杆菌中的高效表达

The expression of hepatitis A virus coding region cDNA in attenuated Salmonella typhimurium

使甲型肝炎病毒基因组ｃＤＮＡ能够在减毒伤寒杆菌中表达，从而通过口服此种减毒伤寒杆菌达到对甲肝免疫的作用。将甲肝病毒编码区ｃＤＮＡ插入高效表达质粒ＰＢＶ２２１，转化大肠杆菌ＤＨ５α，筛选并鉴定阳性克隆。将阳性克隆用电转移法转化减毒的伤寒杆菌ＢＲＤ５０９。用温度诱导法诱导目的基因在伤寒杆菌中表达。通过Ｗｅｓｔｅｒｎ Ｂｌｏｔ和ＥＬＩＳＡ检测表明，表达蛋白可与人抗甲肝ＩｇＧ发生特异性反应。用此伤寒杆菌口服或腹腔注射免疫小鼠，小鼠血清检测到抗甲型肝炎病毒的抗体。结论：甲肝病毒ｃＤＮＡ可以在减毒伤寒杆菌中表达，且表达的蛋白质在体内、外有免疫原性。

Abjective to express the Hepatitis A virus genome cDNA attenuated salmonella Typhimurium and to find the immunity to hepatitis A virus after administrated that salmonella Typhimurium with hepatitis A virus gemome to the mouse(Method)hepatitis A virus coding region cDNA were inserted expressive plasmid PBV221 and then were transform into E coli DH5α.The postive plasmid transforms attenuated salmonella typhimurium BRD509 by electroporation The aim-gene is expressed is Salmonella (Result)That the expressed protein reacted specifically with HAVIgG antibody were demonstrated by ELISA and Western Blot The transformed salmonella were administrated to mice by oral and abdominal,HAVIgG antibody is existed in the mouse serum (Conclusion)Hepatitis A virus cDNA can be expressed in attenuated Salmonella typhimurium and the expressed protein can induce the specific immune response to HAV