【摘要】：為了解云南藍(lán)舌病病毒(Bluetongue virus,BTV)1型M6基因流行株的遺傳變異及其與國(guó)內(nèi)外流行病毒的遺傳進(jìn)化關(guān)系,試驗(yàn)從細(xì)胞培養(yǎng)物中分別提取4株云南分離株BTV-1(Y863、SZ120169、6-12和7-12)RNA,用M6基因特異引物進(jìn)行RT-PCR擴(kuò)增和測(cè)序,采用生物信息學(xué)軟件對(duì)獲得的M6基因編碼區(qū)序列進(jìn)行核苷酸、氨基酸同源性比對(duì)及遺傳進(jìn)化分析。結(jié)果表明,分別獲得4株云南分離株BTV-1 M6基因1 763bp序列;4株云南分離株BTV-1核苷酸同源性在95.2%~99.9%之間,氨基酸同源性在97.6%~99.8%之間,1979年師宗分離的Y863病毒毒株與2012年師宗(SZ120169)、2013年江城(6-12、7-12)分離的3株病毒毒株核苷酸同源性分別為95.5%、95.2%和95.2%,氨基酸同源性分別為97.6%、98.4%和98.2%,而近兩年(2012、2013)分離病毒核苷酸和氨基酸同源性較高,分別在96.9%~99.9%和99.1%~99.8%之間;遺傳進(jìn)化分析發(fā)現(xiàn),4株云南分離株BTV-1為Eastern基因群病毒,它們之間核苷酸和氨基酸同源性分別為95.2%~99.9%和97.6%~99.8%;進(jìn)一步分析發(fā)現(xiàn)4株云南分離株BTV-1與希臘及澳大利亞BTV-1型毒株親緣關(guān)系較近,核苷酸和氨基酸同源性分別為90.4%~95.6%和95.1%~99.1%,而與地中海國(guó)家(意大利、法國(guó)、阿爾及利亞、摩洛哥和突尼斯)和南非毒株關(guān)系較遠(yuǎn),核苷酸和氨基酸同源性分別在83.8%和95.7%以下。4株云南分離株BTV-1屬于Eastern基因群病毒,云南分離株BTV-1 M6基因在自然進(jìn)化中發(fā)生遺傳變異緩慢,該基因可以用來(lái)進(jìn)行BTV-1基因群分布及毒株的地理區(qū)域來(lái)源相關(guān)的研究。
[Abstract]:In order to understand the genetic variation of Yunnan bluetongue virus (Bluetongue virus,BTV) type 1 M6 gene and its relationship with the domestic and foreign epidemic viruses, four Yunnan strain BTV-1 (Y863) were isolated from cell cultures. SZ120169,6-12 and 7-12) RNA, were amplified by M6 gene specific primers and sequenced. Nucleotide, amino acid homology and genetic evolution were analyzed by bioinformatics software. The results showed that BTV-1 M6 gene 1 763bp sequence was obtained from 4 Yunnan isolates. The nucleotide homology and amino acid homology of BTV-1 and amino acid were 95.2% and 99.8%, respectively. The Y863 virus strains isolated in 1979 and 2012 had the same nucleotide identity and amino acid homology (SZ120169), and the nucleotide homology of Y863 virus isolated from Yunnan was 99.9%, and the amino acid homology was 99.8%. The nucleotide homology and amino acid homology of three virus strains isolated from Jiangcheng in 2013 were 95.5% and 95.2%, respectively, and the amino acid homology were 97.6% and 98.2%, respectively. The nucleotide and amino acid homology of virus isolated in the past two years (2012 / 2013) was between 99.9% and 99.8%, respectively, and the homology of nucleotide and amino acid was 99.9% and 99.1%, respectively. The results of genetic evolution analysis showed that BTV-1 was a Eastern gene group virus, and the nucleotide and amino acid homology between them were 95.299.9% and 97.6%, respectively. Further analysis showed that the four Yunnan isolates were closely related to the BTV-1 strains of Greece and Australia. The homology of nucleotide and amino acid were 90.4% and 95.6%, respectively, and 95.1% and 99.1%, respectively. They are more distant from the Mediterranean countries (Italy, France, Algeria, Morocco and Tunisia) and from South Africa. The nucleotide and amino acid homology were below 83.8% and 95.7%, respectively. 4 strains of Yunnan isolate BTV-1 belong to Eastern gene group virus. The genetic variation of BTV-1 M6 gene in Yunnan isolate is slow in natural evolution. This gene can be used to study the distribution of BTV-1 gene group and the geographical source of the virus strain.
【作者單位】： 云南省畜牧獸醫(yī)科學(xué)院云南省熱帶亞熱帶動(dòng)物病毒病重點(diǎn)實(shí)驗(yàn)室;
【基金】：云南省應(yīng)用基礎(chǔ)研究計(jì)劃(青年項(xiàng)目)(2012FD082) 國(guó)家公益性(農(nóng)業(yè))行業(yè)專(zhuān)項(xiàng)(201303035)
【分類(lèi)號(hào)】：S852.65

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