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目的测定2014年云南省狂犬病病毒(RABV)的G基因序列,阐明它们的分子生物学特征。方法用RT-PCR法对RABV的G基因进行全序扩增,并对它们核苷酸序列和推导氨基酸序列进行同源性和进化树分析以及氨基酸位点分析。结果经序列测定,共获得16株RABV的G基因全序列。进化分析表明,本次云南RABV均为基因I型,与四川、广西及既往云南流行株亲缘关系较近,与东南亚流行株也具有较近的亲缘关系。云南株间的核苷酸和氨基酸同源性分别为99.4%~100.0%和99.2%~100.0%,它们与参考株的核苷酸和氨基酸同源性分别86.1%~99.8%和92.9%~99.8%。云南16株RABV的主要抗原性、嗜神经性、毒力及糖基化等功能位点均未发生明显改变,但信号肽氨基酸序列与参考株间差异较多。结论2014年云南RABV间无明显差异,均属于我国主要流行的基因I型中的中国I进化群。云南株RABV的G蛋白重要功能位点未发生变异,仍然保持其特有的抗原性和致病性等特征。
Objective To determine the sequence of G gene of Yunnan rabies virus (RABV) in 2014 and clarify their molecular biological characteristics. Methods The G gene of RABV was amplified by RT-PCR, and their nucleotide and deduced amino acid sequences were analyzed by homology, phylogenetic tree analysis and amino acid sequence analysis. Results After sequencing, a total of 16 RABV G gene sequences were obtained. Phylogenetic analysis showed that RABV in Yunnan was a type I genotype, which had close genetic relationship with epidemic strains in Yunnan, Sichuan and Yunnan in the past, and also had a close genetic relationship with the endemic strains in Southeast Asia. The nucleotide and amino acid homologies of Yunnan strains were 99.4% -100.0% and 99.2% -100.0%, respectively. The nucleotide and amino acid homologies of the two strains were 86.1% -99.8% and 92.9% -99.8 %. The main antigenic, neurotropic, virulent and glycosylated functional sites of 16 RABV strains in Yunnan did not change significantly, but there were significant differences between the amino acid sequences of signal peptides and reference strains. Conclusion There was no significant difference between Yunnan RABV in 2014 and all belong to the Chinese I evolutionary group among the major genotypes I in China. Yunnan Strain RABV G protein important functional site did not mutate, still maintain its unique antigenicity and pathogenicity and other characteristics.