云南蓝舌病病毒1型毒株M6基因序列分析

doi:10.16431/j.cnki.1671-7236.2016.02.008

摘要/Abstract

摘要： 为了解云南蓝舌病病毒(Bluetongue virus,BTV) 1型M6基因流行株的遗传变异及其与国内外流行病毒的遗传进化关系,试验从细胞培养物中分别提取4株云南分离株BTV-1 (Y863、SZ120169、6-12和7-12) RNA,用M6基因特异引物进行RT-PCR扩增和测序,采用生物信息学软件对获得的M6基因编码区序列进行核苷酸、氨基酸同源性比对及遗传进化分析.结果表明,分别获得4株云南分离株BTV-1 M6基因1 763 bp序列;4株云南分离株BTV-1核苷酸同源性在95.2%~99.9%之间,氨基酸同源性在97.6%~99.8%之间,1979年师宗分离的Y863病毒毒株与2012年师宗(SZ120169)、2013年江城(6-12、7-12)分离的3株病毒毒株核苷酸同源性分别为95.5%、95.2%和95.2%,氨基酸同源性分别为97.6%、98.4%和98.2%,而近两年(2012、2013)分离病毒核苷酸和氨基酸同源性较高,分别在96.9%~99.9%和99.1%~99.8%之间;遗传进化分析发现,4株云南分离株BTV-1为Eastern基因群病毒,它们之间核苷酸和氨基酸同源性分别为95.2%~99.9%和97.6%~99.8%;进一步分析发现4株云南分离株BTV-1与希腊及澳大利亚 BTV-1型毒株亲缘关系较近,核苷酸和氨基酸同源性分别为90.4%~95.6%和95.1%~99.1%,而与地中海国家(意大利、法国、阿尔及利亚、摩洛哥和突尼斯)和南非毒株关系较远,核苷酸和氨基酸同源性分别在83.8%和95.7%以下.4株云南分离株BTV-1属于Eastern基因群病毒,云南分离株BTV-1 M6基因在自然进化中发生遗传变异缓慢,该基因可以用来进行BTV-1基因群分布及毒株的地理区域来源相关的研究.

关键词: 蓝舌病病毒; M6基因; 序列分析

Abstract: The object of this study was to investigate the evolution and variation of NS1 encoding gene (M6) of Bluetongue virus 1 (BTV-1) from Yunnan province and the evolutionary relationship of strains which from Yunnan province and other countries.RNA were extracted from four strains (Y863,SZ120169,6-12 and 7-12),and M6 gene were amplified by using specific primer and sequenced,which were analyzed by using bioinformatics software for nucleotides homology and phylogenetic relationships.The results showed that four strains M6 gene were 1 763 bp;The homology of nucleotides among four strains M6 gene were 95.2% to 99.9%,the amino acids among four strains M6 gene were 97.6% to 99.8%,the homology of nucleotides between Y863 (1979) and 3 strains (SZ120169,6-12 and 7-12) were 95.5%,95.2% and 95.2%,the amino acids between Y863 (1979) and 3 strains were 97.6%,98.4% and 98.2%,the homology of nucleotides and amino acids were high (96.9% to 99.9% and 99.1% to 99.8%,respectively) among four Yunnan strains.BTV was divided into two clusters (Western and Eastern) and four strains (BTV-1) from Yunan province belong to Eastern cluster.The homology of nucleotides and amino acids among four Yunnan strains was 95.2% to 99.9% and 97.6% to 99.8% respectively;The genetic distance were close among four Yunnan strains and strains from Greece and Australia,the homology of nucleotides and amino acids between them were 90.4% to 95.6% and 95.1% to 99.1%;The genetic distance were distinct among four Yunnan strains and strains from Mediterranean countries (Italy,Fance,Algeria,Morocco and Tunisia) and South Africa;The homology of nucleotides and amino acids between them were below 83.8% and 95.7%,so we found that gene clusters distribution was related to the geographical distribution for BTV.In a conclusion,four Yunnan strains belong to Eastern cluster and the speed of genetic variation of M6 from Yunnan province was slow,so M6 gene could be used in study of gene group of distribution and origin of geographical area.

Key words: BTV; M6 gene; sequence analysis

中图分类号:

S852.65⁺9.4

李楠, 朱建波, 肖雷, 李乐, 杨恒, 孟锦昕, 何于雯, 李华春. 云南蓝舌病病毒1型毒株M6基因序列分析[J]. , 2016, 43(2): 340-347.

LI Nan, ZHU Jian-bo, XIAO Lei, LI Le, YANG Heng, MENG Jin-xin, HE Yu-wen, LI Hua-chun. Sequence Analysis of Bluetongue Virus 1 M6 Gene in Yunnan Province[J]. , 2016, 43(2): 340-347.

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