小反刍兽疫病毒非结构蛋白V基因的克隆及其编码蛋白的结构与功能

采用RT-PCR技术从小反刍兽疫病毒总RNA中扩增到P基因一段896bp序列,克隆至pET-30a(+)载体中,对其进行最大开放阅读框691位定点突变(插入一个G碱基),获得小反刍兽疫病毒V基因,利用生物信息学软件对麻疹病毒属V蛋白氨基酸序列进行比对,同时使用I-TASSER服务器对小反刍兽疫病毒V蛋白三级结构进行同源建模,分析其结构与功能。结果显示,小反刍兽疫病毒V蛋白氨基酸序列与麻疹病毒、犬瘟热病毒、牛瘟病毒和海豹瘟病毒V蛋白的相似性分别为61.1%、56.0%、61.7%和51.7%,三级结构预测显示V蛋白由多α-螺旋的N-端结构域、中间结构域和含锌指结构的C-端结构域组成。本研究成功克隆了小反刍兽疫病毒V基因,预测分析了V蛋白结构和功能的关系,为其分子生物学和功能的进一步研究奠定了基础。

Cloning of the nonstructural protein V gene of peste des petits ruminants and analysis of the structure and function of the protein encoded by V gene

A 896 bp fragment of the P gene was amplified from peste des petits ruminants virus(PPRV) total RNA by RT-PCR and cloned into pET-30a(+) vector.V gene was obtained by site-directed mutagenesis in ORF 691 site(insert a G).V protein amino acid sequences of Morbillivirus were aligned,the three-dimensional structure of the protein was developed by homology modeling methods using I-TASSER.Sequence comparison revealed that the homologies of deduced amino acid sequences between PPRV and measles virus,canine distem...