含伪狂犬病毒完整UL49基因及部分UL48基因片段的克隆与序列分析  

Cloning and Sequences Determination of the Fragment Containing UL49 Gene and Partial UL48 Gene of Pseudorabies Virus(PRV)

作　　者：方六荣[1] 牛传双[1] 肖少波[1] 余晓岚[1] 陈桦[1] 陈焕春[1] 

机构地区：[1]华中农业大学畜牧兽医学院,武汉430070

出　　处：《华中农业大学学报》2002年第4期306-310,共5页Journal of Huazhong Agricultural University

基　　金：湖北省自然科学基金 (2 0 0 0J10 0 )资助项目;华中农业大学大学生科技创新基金 (SRF 2 0 0 1A3 0 )资助项目

摘　　要：以地高辛标记的伪狂犬病毒 (PRV)Ea株UL4 9.5基因片段为探针 ,同SalI、KpnI、BamHI消化的PRVEa株基因组DNA进行Southern杂交 ,确定SalI 2 .8kb、KpnI 17.0kb、BamHI 2 5 .0kb片段中含UL4 9基因。将SalI2 .8kb片段克隆到pUC119中 ,利用其中的BamHI进行亚克隆 ,获得重组质粒pUC2 .0。测定约 2 .0kb片段的全序列并进行序列分析 ,发现该片段包含UL5 0基因部分编码区 ,UL4 9.5基因和UL4 9基因的完整编码区 ,并且在UL4 9基因下游 ,还存在一段约 36 9个碱基的序列 ,经与人单纯疱疹病毒I型 (HSV 1)、牛单纯疱疹病毒I型 (BHV 1)的UL4 8基因进行比对 ,具有较高的同源性 ,推测为PRVUL4 8基因的部分编码区。The genomic DNA of Pseudorabies virus(PRV) strain Ea was digested with Sal I, Kpn I, Bam HI and hybridized with the Dig labbled DNA fragment containing UL 49.5 gene of PRV. The Southern blot shows that the Sal I 2.8 kb, Kpn I 17.0 kb and Bam HI 25.0 kb contain complete UL 49 gene. The Sal I 2.8 kb fragment was cloned into vector pUC119 and a 2.0 kb fragment was further sub cloned to yield the recombinant plasmid pUC2.0.The complete sequence of the 2.0 kb fragment was determined by Sanger's sequencing technique. The sequences analysis shows that the 2.0 kb fragment is comprised of partial UL 50 gene, complete UL 49.5 and UL 49 genes. It is interesting that there exist additional 369 bases downstream the poly(A) signal sequence of UL 49 gene. After comparison with HSV 1 and BHV 1, the sequences are found to have significant similarity to the UL 48 gene of HSV 1 and BHV 1, which indicates that the additional sequences are the potential partial UL 48 gene of PRV.

关 键 词：基因克隆 伪狂犬病毒 完整UL49基因 部分UL48基因 序列分析 

分 类 号：S852.659.1[农业科学—基础兽医学]