论文部分内容阅读
为解析牛副流感病毒3型(BPIV3)SD2014分离株P蛋白磷酸化位点与功能性结构域,首先对P基因全长进行克隆测序,并与SD0835株P基因进行比较;再利用DNAStar中的MegAlign软件,以Clustal W方法比较了副黏病毒不同种属15株参考毒株的P蛋白氨基酸序列的同源关系,并对23株牛、人和猪副流感病毒3型P蛋白氨基酸序列进行比对筛选保守的氨基酸位点;运用在线Netphos 2.0Server和NetPhosK 1.0Server软件分别预测BPIV3SD2014毒株P蛋白潜在的磷酸化位点和特异性磷酸化蛋白激酶作用位点,最后将P蛋白4个主要功能性结构域氨基酸位置与潜在磷酸化位点一一对应。结果发现,SD2014毒株P基因核苷酸序列与SD0835株同源性为99%,P蛋白氨基酸序列与猪副流感病毒3型(SPIV3)同源性最高为73.2%。P蛋白在398~479aa含有1段螺旋卷曲结构,推测是介导P蛋白四聚体的形成以及与L蛋白的相互作用的位点,同时发现PKC是唯一一个贯穿于所有功能结构域的蛋白激酶。本研究为进一步解析P蛋白磷酸化在调控BPIV3转录与复制过程中的作用奠定基础,同时也为研究P蛋白的生物学功能提供支撑。
In order to resolve the P protein phosphorylation site and functional domain of BPIV3 SDV-B2014 isolate, the full-length P gene was first cloned and sequenced and compared with the P gene of SD0835 strain. MegAlign software was used to compare the homology of the P protein amino acid sequences of 15 reference strains of different species of paramyxovirus by the Clustal W method and to compare the amino acid sequence of the 23 protein of P, To screen the conservative amino acid sites; using online Netphos 2.0Server and NetPhosK 1.0Server software to predict potential P protein phosphorylation sites and specific phosphorylation protein kinase site of BPIV3SD2014 strain, respectively, and finally the four major functions of P protein The amino acid positions of the sex domains correspond to the potential phosphorylation sites. The results showed that the nucleotide sequence of P gene of SD2014 strain shared 99% homology with SD0835 and the highest homology of P protein with that of SPIV3 was 73.2%. The P protein contains a 1-segment helix-coil structure at 398-479aa. It is presumed to be a site that mediates the formation of tetrameric P and the interaction with L protein. PKC is also found to be the only protein kinase that permeates all functional domains . This study laid the foundation for further analysis of the role of P-protein phosphorylation in regulating the transcription and replication of BPIV3, and also provided support for studying the biological function of P protein.