【摘要】 提取伪狂犬病病毒 (PRV )Bartha K61株基因组DNA ,用限制性内切酶KpnⅠ充分消化 ,回收 5 .9× 10 3 b片段(J片段 ) ,将其克隆于质粒 pUC119KpnⅠ位点上 ,获得pBKJ。用一对针对PRVTK基因的特异性引物对重组质粒进行PCR鉴定 ,证明其中含有PRVTK基因。然后用KpnⅠ、PstⅠ和BamHⅠ等限制性内切酶对其进行酶切分析 ,确定了克隆片段的物理图谱。进一步研究证实TK基因位于其中的KpnⅠ BamHⅠ或KpnⅠ PstⅠ片段中。然后亚克隆此KpnⅠ PstⅠ片段 ,并进行了序列测定。更多还原

【Abstract】 The genome DNA extracted from pseudorabies virus (PRV) Bartha k61 strain was sufficiently digested by restriction endonuclease Kpn I, from which the 5.9×10 3 basepair fragment (J fragment) was obtained and cloned into plasmid pUC 119 Kpn I site and the pBK J was got. The recombinant plasmid was identified using a pair of primers specific to PRV TK gene, which proved that the PRV TK gene just was in the recombinant plasmid. Then the recombinant plasmid was cut and analysed with the restriction endonuclease Kpn I, Pst I and BamH I and the physical map of the cloned fragment was determined, which further confirmed that the TK gene was in the Kpn I BamH I fragment or in the Kpn I Pst I fragment, The Kpn I Pst I fragment was subcloned and sequenced, which is favourable for improving and applying the alive carrier of PRV.更多还原