马立克氏病病毒Rispens株和RL株B抗原基因的分离、克隆及初步酶切分析  

Amplification, Cloning and Restriction Enzyme Analysis of Glycoprotein B Antigen Genes of Marek′s Disease Viruses (MDVs) Rispens strain and RL Strain

作　　者：邢力[1] 彭大新[1] 刘秀梵[1] 张如宽[1] 吴艳涛[1] 

机构地区：[1]扬州大学农学院动物医学系

出　　处：《中国兽医学报》1998年第4期317-320,共4页Chinese Journal of Veterinary Science

基　　金：江苏省科委应用基础研究项目

摘　　要：将马立克氏病病毒（ＭＤＶ）血清Ⅰ型Ｒｉｓｐｅｎｓ株和ＲＬ株分别接种于原代鸡胚成纤维细胞，待出现８０％以上细胞病变后收获，经蛋白酶Ｋ消化，酚、氯仿抽提，乙醇沉淀细胞和病毒的总ＤＮＡ（ｔｏｔａｌＤＮＡ）。以此为模板，根据已发表的Ｂ抗原基因核苷酸序列设计１对引物，通过聚合酶链式反应（ＰＣＲ）扩增出１条约２．８５ｋｂ的特异性条带，斑点杂交证实其为Ｂ抗原基因。双酶切消化后，克隆到质粒ｐＵＣ１９中。酶切分析表明，在这２株病毒的Ｂ抗原基因上，ＨｉｎｄⅢ、ＥｃｏＲＶ、ＢａｍＨＩ、ＥｃｏＲＩ的酶切位点分布与超强毒ＲＢＩＢ株、标准强毒ＧＡ株的完全相同。Purified DNAs from chicken embryo fibroblast cultures infected respectively with rispens strain and RL strain of Marek′s disease virus (MDV) were used as templates. The specific fragments of about 2.85 kb were respectively amplified through polymerase chain reaction and shown to be the genes of glycoprotein B (gB) of rispens strain and RL strain respectively after chemiluminescence dot blot hybridization with a digoxigeninlabelled MDV gB gene specific oligonucleotide probe. It has been demonstrated that Hind Ⅲ, Eco RV, Bam HI, Eco RI restriction enzyme cleavage patterns of the gBs of rispens strain and RL strain were identical to those of RBIB strain and GA strain.

关 键 词：马立克氏病病毒 B抗原 限制性酶切分析 PCR 

分 类 号：S852.65[农业科学—基础兽医学] Q78[农业科学—兽医学]