水貂阿留申病毒VP2基因高变区的多变性分析
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摘要
为了了解青岛地区水貂阿留申病毒(ADV)VP2基因高变区的变异情况,试验参考基因库中ADV VP2基因设计1对引物,利用PCR技术扩增VP2基因高变区序列,应用DNAMAN进行序列分析和同源性比较。结果表明:扩增的6株ADV VP2基因高变区碱基长度均为226 bp,与预期目的条带大小一致。6株ADV VP2基因高变区核苷酸同源性为92.9%~100%,与已知国内外12株ADV VP2基因高变区相比核苷酸同源性为86.7%~97.3%。其中除2株基因序列完全相同外,其他4株都有很大的突变,与国内外毒株也有较大的差异,表明ADV VP2基因具有多变性。
The aim of the present study was to understand the variation of VP2 gene hypervariable region in mink aleutian virus( ADV) in Qingdao. A pair of primers were designed according to the nucleotide sequence of ADV VP2 gene in Genbank. The hypervariable region gene of VP2 was amplified by PCR from ADV genome. Sequence analysis and homology comparison were performed by using the software DNAMAN. The results showed that the length of the amplified six ADV VP2 genes was 226 bp,which was consistent with the intended purpose strip size. Sequence analysis showed that the nucleotide homology of the high variable region of the ADV VP2 gene in the six strains was 92. 9% ~ 100%. The nucleotide homology was 86. 7% ~ 97. 3% compared with the ADV VP2 gene of the domestic and foreign 12 strains. Among them,except 2 strains of the same gene sequence,there were very large mutation in the other 4 strains. And there were also large difference compared with the domestic and foreign strains. The results indicated that the ADV VP2 gene was variable.
The aim of the present study was to understand the variation of VP2 gene hypervariable region in mink aleutian virus( ADV) in Qingdao. A pair of primers were designed according to the nucleotide sequence of ADV VP2 gene in Genbank. The hypervariable region gene of VP2 was amplified by PCR from ADV genome. Sequence analysis and homology comparison were performed by using the software DNAMAN. The results showed that the length of the amplified six ADV VP2 genes was 226 bp,which was consistent with the intended purpose strip size. Sequence analysis showed that the nucleotide homology of the high variable region of the ADV VP2 gene in the six strains was 92. 9% ~ 100%. The nucleotide homology was 86. 7% ~ 97. 3% compared with the ADV VP2 gene of the domestic and foreign 12 strains. Among them,except 2 strains of the same gene sequence,there were very large mutation in the other 4 strains. And there were also large difference compared with the domestic and foreign strains. The results indicated that the ADV VP2 gene was variable.
引文
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[7]张瑞,李艳伍,许兰菊,等.阿留申病毒强致病性毒株非结构蛋白基因的克隆及序列分析[J].中国动物检疫,2010,27(2):27-31.
[8]田国宁,张金玲,李凯,等.水貂阿留申病毒环介导等温扩增(LAMP)检测方法的建立[J].中国动物检疫,2015,32(6):79-82.
[9]张蕾,胡博,白雪,等.一株水貂阿留申病毒VP2基因的高变区测序分析[J].病毒学报,2015,31(3):226-230.
[10]刘立兵,冯海峰,石坤,等.水貂阿留申病毒部分VP2基因的原核表达及免疫学分析[J].经济动物学报,2013,71(1):12-14,18.
[11]张蕾,马凡舒,孙彦刚,等.水貂阿留申病诊断技术的研究进展[J].安徽农业科学,2015,43(6):156-157,160.
[12]邵西群,章秀婷,岳志刚,等.水貂血液阿留申病病毒及其抗体的检测与分析[J].中国兽医学报,2014,34(12):1893-1898,1905.
[13]李艳伍,张瑞,姜平,等.水貂阿留申病病毒分子生物学研究进展[J].动物医学进展,2009,30(9):100-104.
[2]WANG Z J,WU W,HU B,et al.Molecular epidemiology of Aleutian mink disease virus in China[J].Vrius Res,2014,184(3):14-19.
[3]WU W H,BLOOM M E,BERRY B D,et al.Expression of Aleutian mink disease parvovirus capid proteins in a baculovirus expression system for potential diagnostic use[J].J Vet Diagn Invest 1994,6(1):23-29.
[4]CASTELRUIZ Y,BLIXENKVONE-MOLLER M,AASTED B.DNA vaccination with the Aleutian mink disease virus NS1 gene confers partial protection against disease[J].Vaccine,2005,23(10):1225-1231.
[5]梁冬莹,华育平,曾祥伟.水貂阿留申病毒VP2基因主要抗原表位区的原核表达[J].东北林业大学学报,2007,6(6):39-41.
[6]刘东旭,李健明,时坤,等.水貂阿留申病毒5个分离株的全基因测序及遗传变异分析[J].动物医学进展,2015,36(8):35-39.
[7]张瑞,李艳伍,许兰菊,等.阿留申病毒强致病性毒株非结构蛋白基因的克隆及序列分析[J].中国动物检疫,2010,27(2):27-31.
[8]田国宁,张金玲,李凯,等.水貂阿留申病毒环介导等温扩增(LAMP)检测方法的建立[J].中国动物检疫,2015,32(6):79-82.
[9]张蕾,胡博,白雪,等.一株水貂阿留申病毒VP2基因的高变区测序分析[J].病毒学报,2015,31(3):226-230.
[10]刘立兵,冯海峰,石坤,等.水貂阿留申病毒部分VP2基因的原核表达及免疫学分析[J].经济动物学报,2013,71(1):12-14,18.
[11]张蕾,马凡舒,孙彦刚,等.水貂阿留申病诊断技术的研究进展[J].安徽农业科学,2015,43(6):156-157,160.
[12]邵西群,章秀婷,岳志刚,等.水貂血液阿留申病病毒及其抗体的检测与分析[J].中国兽医学报,2014,34(12):1893-1898,1905.
[13]李艳伍,张瑞,姜平,等.水貂阿留申病病毒分子生物学研究进展[J].动物医学进展,2009,30(9):100-104.