摘要
利用甲型肝炎病毒cDNA,将编码病毒主要结构蛋白基因的不同区段插入到pUC19和pUR290(或pUR292),组建成一系列表达质粒,使甲肝病毒结构蛋白与β-半乳糖苷酶形成融合蛋白,在大肠杆菌中得到表达。表达克隆pHABB-290含部分VP3的羟基端及VP1氨基端的一半;pHABP-290含VP1的中间一段;pHABSR1-19含小部分VP3和近完整的VP1;pHAPH292含VP2羟基端的约2/3和VP3的绝大部份。用聚丙烯酰胺凝胶电泳分析上述克隆,蛋白染色后,在预计位置均可见到很浓的表达蛋白带,而且表达的融合蛋白均受IPTG诱导。Westernblot分析发现,只有pHABSR1-19表达的融合蛋白可与甲肝病人恢复期血清起免疫反应。这说明表达的融合蛋白含有甲肝病毒自然感染时所具有的抗原性。
Using the cDNA of hepatitis A virus ( HAV ) and expression plasmids of pUR 292 (or pUR290) and pUC19, four recombinant plasmids expressing the major structure proteins of HAV were constructed.After transformation into E.coli and IPTG induction, VP1,VP2 and VP3 were successfully expressed in abundant quantity as fusion proteins with β-galacto-sidase. All the expressed fusion proteins appeared at expected position on SDS-PAGE analysis. Western-blot analysis showed that only the recom-binant fusion protein containing nearly complete VP1 protein could react with human HAV convalescence sera,suggesting that the E coli expressed VPl protein possesses the antigenicity of HAV.
出处
《病毒学报》
CAS
CSCD
北大核心
1989年第4期312-317,共6页
Chinese Journal of Virology
关键词
甲肝病毒
外壳蛋白
基因表达
Hepatitis A virus Capsid protein Gene expression
