马立克病病毒疫苗株CVI988的38kd磷蛋白基因双氨基酸突变株  

CONSTRUCTION OF A DOUBLE-AMINO ACID MUTANT IN 38kd PHOSPHORYLATED PROTEIN OF MAREK'S DISEASE VIRUS VACCINE STRAIN CVI988

作　　者：崔治中[1] 秦爱建[2] LeeL.F. 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018 [2]扬州大学畜牧兽医学院,江苏扬州225009 [3]AvianDiseaseandOncologyLab

出　　处：《山东农业大学学报（自然科学版）》2000年第3期231-235,共5页Journal of Shandong Agricultural University：Natural Science Edition

基　　金：国家"8 6 3"项目!抗鸡马立克病多价基因工程疫苗研究 (1 0 1- 0 5- 0 3- 0 2 )资助项目

摘　　要：利用单克隆抗体H1 9及间接免疫荧光抗体反应 (IFA) ,从经马立克病病毒 (MDV)强毒株Md1 1的38kd磷蛋白 (pp38)基因克隆质粒DNA转染的MDV疫苗毒CVI988株感染的鸡胚成纤维细胞中 ,筛选到一个点突变株CVI/rpp38(AA)。DNA序列测定结果表明 ,该点突变株pp38基因ORF的第 32 0位和 32 6位碱基均已从亲本CVI988株的“G”突变为“A” ,导致第 1 0 7和 1 0 9位氨基酸分别从精氨酸和甘氨酸转变为谷氨酰胺和谷氨酸。这个双氨基酸点突变株在IFA和免疫沉淀反应中与pp38特异性单抗H1 9都呈现阳性反应 ,但与另一个 pp38特异性单抗T6 5都完全不反应 ,表现出与亲本病毒完全相反的结果。本研究发现了MDV的pp38上二个特异性不同的抗原表位 ,并以确凿的证据证明了第 1 0 7及 1 0By using indirect fluorescence antibody test (IFA) with pp38-specific monoclonal antibody(Mab) H19,one more point mutant CVI/rpp38(AA) of Marek's disease virus(MDV)vaccine strain CVI988 was screened from the CVI988-infected chicken embryo fibroblast monolayer transfected with virulent MDV pp38 gene plsmid clone DNA.Sequencing data indicated that both bases at #320 and #326 in pp38 ORF of the mutant were switched into bases“A”from“G”of its parent virus CVI988,leading the amino acids at #107 and #109 changed into glutamine and glutamic acid respectively from arginine and glycine of native CVI988.The double-aa mutant was reactive with Mab H19 but not with another pp38-specific Mab T65 in both IFA and immunoprecipitation in contrast to the native CVI988 which was just opposite in reaction with H19 and T65.The study demonstrated the two different epitopes on the pp38 molecule and provided a convincing evidence that the glutamine at #107 and glycine at #109 were critical to the two epitopes H19 and T65 with different specificity.

关 键 词：点突变 特异性 抗原表位 CV 单抗 病毒疫苗 氨基酸 马立克病 MDV 亲本 

分 类 号：S858[农业科学—临床兽医学] S852[农业科学—兽医学]