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根据小反刍兽疫病毒(PPRV)参考序列设计了11对特异性引物,然后用RT-PCR方法分段扩增出PPRV富阳分离株(PPRV-FY)的全基因组序列,用DNAstar软件和Mega软件对PPRV-FY株及参考毒株的基因序列进行了比对和分析,并根据N基因核苷酸序列绘制了系统进化树。结果,PPRV-FY株全长15 948 bp,编码6种结构蛋白(N、P、L、M、F和H),2种非结构蛋白(C和V),PPRV各毒株转录起始序列和转录终止序列都高度保守。同源性分析结果表明,PPRV-FY株与PPRV-Coted′Ⅰvoire89同源性最低,为89.0%,与PPRV/Tibet/30/2007株和PPRV/Tibet/2007株的同源性最高,为97.3%,其基因间隔区N-P长度一致,相似度最高,而M-F相似度最低。通过比较PPRV-FY株与参考毒株基因组末端调控序列,可以看出整个麻疹病毒属GP区和AGP区保守性较高,且存在不同程度的互补。氨基酸序列分析结果表明,PPRV-FY株在保守区域出现21处氨基酸突变,这些突变对其所编码蛋白的结构和功能有何影响尚需要进一步研究。此外,本文还基于N基因序列,对PPRV-FY株及参考毒株进行了系统进化分析。结果表明,PPRV-FY株与其它亚洲源PPRV野毒株共同形成一个拓扑群,属于第IV谱系。
According to the reference sequence of PPRV, 11 pairs of specific primers were designed, and then the complete genome sequence of PPRV-FY was amplified by RT-PCR. DNAstar software and Mega software PPRV-FY strain and reference strain were sequenced and analyzed, and phylogenetic tree was drawn based on the nucleotide sequence of N gene. As a result, the PPRV-FY strain was 15 948 bp in length encoding six structural proteins (N, P, L, M, F and H), two non-structural proteins (C and V) And transcription termination sequences are highly conserved. Homology analysis showed that the homology of PPRV-FY strain with PPRV-Coted’Ⅰvoire89 was the lowest (89.0%), the highest homology with PPRV / Tibet / 30/2007 and PPRV / Tibet / 2007 was 97.3 %, The length of the intergenic region NPs is the same, the highest similarity and the lowest MF similarity. By comparing the regulatory sequences of PPRV-FY strain with those of reference strains, it can be seen that the GP and AGP regions of the entire measles virus are highly conserved and complement each other to varying degrees. The results of amino acid sequence analysis showed that there are 21 amino acid mutations in the conserved region of PPRV-FY strains. The impact of these mutations on the structure and function of the encoded proteins needs further study. In addition, phylogenetic analysis of PPRV-FY strain and reference strain based on N gene sequence was also carried out. The results showed that PPRV-FY strain and other Asian PPRV wild-type strains together form a topological group, belonging to the IV lineage.