表达强毒pp38决定簇的马立克病病毒疫苗毒CVI988点突变株的构建和特性  

CONSTRUCTION AND CHARACTERIZATION OF POINT-MUTATED MAREK'S DISEASE VIRUS VACCINE STRAIN CVI988 EXPRESSING THE VIRULENT EPITOPE ON ITS 38kD PHOSPHORYLATED PROTEIN

作　　者：崔治中[1,2] Le.LF 

机构地区：[1]扬州大学动医系 [2]美国农业部禽病和肿瘤学研究所

出　　处：《病毒学报》1999年第2期147-153,共7页Chinese Journal of Virology

基　　金：国家自然科学基金

摘　　要：用鸡马立克病病毒（ＭＤＶ）强毒ＧＡ株的３８ｋＤ磷蛋白（ｐｐ３８）基因克隆ＤＮＡ转染Ⅰ型弱毒疫苗ＣＡＩ９８８／Ｒｉｓｐｅｎｓ株ＭＤＶ感染的鸡胚成纤维细胞，再用能识别Ⅰ型强毒ｐｐ３８的单克隆抗体Ｈ１９做免疫荧光试验，筛选到能在ｐｐ３８基因上表达强毒株特异性抗原决定簇的定向点突变弱毒株ＣＶＩ／ｒｐｐ３８。用３５Ｓ－蛋氨酸标记的细胞裂解物做免疫沉淀反应表明，单抗Ｈ１９不能识别天然ＣＶＩ９８８株ＭＤＶ中的ｐｐ３８，但却能从感染了重组病毒ＣＶＩ／ｒｐｐ３８的细胞识别出ＭＤＶｐｐ３８蛋白带，如同从ＭＤＶ的其它强毒株识别出ｐｐ３８一样。接种了ＣＶＩ／ｒｐｐ３８病毒的鸡，其ＭＤＶ特异性抗体滴度显著低于接种了原疫苗毒ＣＶＩ９８８／Ｒｉｓｐｅｎｓ克隆株的鸡，这进一步证明了强毒ＭＤＶ的ｐｐ３８具有免疫抑制作用。Chicken embryo fibroblast (CEF) monolayer infected with serotype I Marek's disease virus (MDV) vaccine strain CVI988 were transfected with virulent MDV pp38 gene clone plasmid DNA in lipofect AMI NE solution. A point-mutated recombinant CVI/rpp38, which was expressing a virulent epitope on pp38, was screened and cloned by indirect fluorescent antibody test with monoclonal antibody H19. Immunoprecipitation indicated that H19 could not recognize pp38 complex from native CVI988 strain but did recognize the pp38 complex from the recombinant mutant CVI/rpp38 as from other serotype I virulent MDV strains. Chickens inoculated with the recombinant CVI/rpp38 gave significant lower MDV-specific antibody titers than chickens inoculated with the native CVI988 strain virus did. It demonstrated again that the pp38 from virulent MDV may have immunodepressive effects.

关 键 词：马立克病病毒 38kD磷蛋白 点突变 MDV 

分 类 号：S858.215.3[农业科学—临床兽医学] S852.65[农业科学—兽医学]