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.
出处
《中国兽医学报》
CAS
CSCD
北大核心
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
关键词
马立克氏病病毒
B抗原
限制性酶切分析
PCR
Marek's disease virus
glycoprotein B antigen
polymerase chain reaction
restriction enzyme analysis
