轮状病毒VP7基因在大肠杆菌中的表达及其免疫原性

Expression in Escherichia coli and Immunogenicity of Rotavirus VP7

轮状病毒是世界范围内引起婴幼儿病毒性腹泻的主要病原体。VP7是轮状病毒的主要外壳蛋白和中和抗原 ,是发展基因工程疫苗的首选。把包含全部 3个主要抗原性区域的轮状病毒SA11VP7基因片段以谷胱甘肽S 转移酶融合蛋白的形式在大肠杆菌中进行表达 ,表达产物占菌体总蛋白的 30 %左右。经一步GlutathioneSepharose4B亲和纯化 ,重组蛋白纯度超过 90 %。Westernblot实验表明 ,重组蛋白可被抗SA11的多抗特异地识别。动物实验表明 ,重组抗原可在小鼠和家兔体内诱导VP7特异的抗体和一定水平的SA11中和抗体。

Rotaviruses are the single most important cause of severe diarrhea in young children all over the world.VP7 is the major outer capsid and is a primary candidate for inclusion in a subunit or recombinant vaccine.Part of the VP7 gene containing all the three antigenic regions was expressed as a chimeric protein with glutathione S\|transferase (GST) in \%E.coli.\%The chimeric protein,representing about 30% of the total protein of the recombinant\|plasmid\|carrying bacteria,reacted with polyclonal antibodies raised against whole virus.Immunization of sero\|negative rabbits and mice with purified fusion\|protein generated both virus\|binding and neutralizing antibodies.