【摘要】 将马立克氏病病毒（ＭＤＶ）血清Ｉ型Ｒｉｓｐｅｎｓ株接种原代鸡胚成纤维细胞（ＣＥＦ），待出现８０％以上细胞病变后收获，提取细胞和病毒的总ＤＮＡ．以此为模板，根据Ｂ抗原基因核苷酸序列设计一对引物，通过聚合酶链反应（ＰＣＲ）扩增出一条约２．８５ｋｂ的特异性条带，双酶消化后克隆至质粒ｐＵＣ１９，酶切分析表明该病毒的Ｂ抗原基因上ＨｉｎｄⅢ、ＥｃｏＲⅤ、ＢａｍＨⅠ、ＥｃｏＲⅠ的酶切位点分布与ＲＢ１Ｂ株、ＧＡ株的完全相同，进一步序列分析表明，其核苷酸序列与其他ＭＤＶ血清Ｉ型的ｇＢ基因完全一致更多还原

【Abstract】 DNA from chicken embryo fibroblast (CEF) cultures infected with Rispens strain of Marek′s disease virus (MDV) was used as templates and primers was designed on basis of RB 1B strain. The specific fragment with the size of about 2.85 kb was successfully amplified through polymerase chain reaction (PCR) and cloned into pUC19. It has been demostrated that HindⅢ, EcoRⅤ, BamHⅠ and EcoRⅠ restriction enzyme cleavage patterns and the nucleotide sequence of gB from Rispens was identical to those of RB 1B strain and GA strain.更多还原