三株广西狂犬病病毒NS基因和M基因的克隆与序列分析

Cloning and sequence analysis of NS and M gene of rabies virus isolated from Guangxi province

本研究设计了一对特异性引物NSM1/NSM2,对三株广西狂犬病病毒NS和M基因同时进行了RT-PCR扩增、克隆和测序.同源性分析表明,三株广西野毒NS基因核苷酸同源性为87.2%～98.4%,M基因核苷酸同源性为90.1%～99.7%;与固定毒和狂犬病相关病毒比较,NS基因分别为79.9%～82.8%和69.7%～71.0%;M基因的分别为82.8%～87.8%和75.0%～77.8%.三株野毒NS基因氨基酸同源性为93.3%～98.7%,M基因氨基酸同源性分别为97.5%～100%.表明广西各地毒株之间亲缘关系不同,但最为相近;与狂犬病固定毒株亲缘关系较远;与狂犬病相关病毒亲缘关系最远.

A pair of primers NSM1/NSM2 were designed and synthesized according to the reported rabies virus cDNA sequences in GenBank, and the cDNA fragments including phosphoprotein（NS） gene and matrix protein（M） gene of three rabies virus strains isolated from Guangxi were amplified by reverse transcription-polymerase chain reaction （RT-PCR）, respectively. The cDNA fragments were cloned and then sequenced. The nucleotide sequence of NS and M gene and their deduced amino acid sequence were compared with that of other rabies virus strains and rabies-related virus strains published previously. The nucleotide homology of NS gene among three rabies virus strains isolated from Guangxi was 87.2 % to 98.4 % and in M gene among these three street strains was 90.1% to 99.7 % ; the nucleotide homology between rabies virus strains from Guangxi and from fixed strains or rabies - related virus were ranged from 79.9% to 82.8% or from 69.7 % to 71.0 % in NS gene,and from 82.8 % to 87.8 % or from 75.0 % to 77.8 % in M gene, respectively. The homology of deduced amino acid sequence of NS gene among the three strains was 93.3 % to 98.7 %, and that of among M gene was 97.5 % to 100 %. These results suggested that genetic relationship in the three strains isolated from different place of Guangxi differ but is rather close and there is a big difference between strains from Guangxi and fixed rabies virus strains and rabies-related strains.