马立克病病毒群共同性抗原P79蛋白质基因的鉴定和定位

IDENTIFICATION AND LOCALIZATION OF THE MAREK'S DISEASE VIRUS GROUP-COMMON ANTIGEN P79 GENE

马立克病病毒(MDV)的79kD蛋白质是该属3个型共有的抗原。用抗该抗原的单克隆抗体T81(MAb T81),从建于大肠杆菌噬菌体λgt11中MDV感染的鸭胚成纤维细胞基因组DNA的基因库中,筛选到一个能表达该蛋白质片段的克隆GB-2。对该重组噬菌体的DNA做酶切分析,用PCR扩增插入片段,结果表明GB-2含一个2.5kb来自MDV的插入片段。用标记的该片段DNA作探针,分别与GA株MDV感染的鸭胚成纤维细胞基因组DNA的限制性内切酶消化物及MDV基因组DNA的BamH I-B片段做Southern blot和分子杂交反应,结果证明,编码MDV属共同抗原79kD蛋白质的基因位于其基因组DNA酶切图的BamH I-R片段及B和O片段与R联结的部分。

Marek's disease virus ( MDV ) 79kD protein ( P79 ) is an antigen com-mon for 3 serotypes of the virus group. By using monoclonal antibody T81 against MDV, a P79-expression recombinant phage clone GB-2 was isolated from the MDV-infected duck embryo fibroblast ( DEF ) cell ge-nomic DNA library which was built in the expression vector E.coli lam-bda gtll phage. Restriction enzyme analysis and polymerase chain reac-tion indicated that the GB-2 clone contained a MDV-origin insert of 2.5kb. As theprobe,the labeled GB-2 clone was hybridized withGA strain MDV-infected DEF genomic DNA digested by different restriction en-zymes and the knowh BamHI-B fragment of MDV genomic DNA in Southern blot. The results demonstrated that the gene encoding the MDV group-common antigen P79 is located in the whole BamHI-R fragment and parts of O and B fragments which were connected with the R fragment on the MDV genomic DNA map.