樱桃谷鸭源鹅出血性多瘤病毒VP3基因的克隆与序列分析

发布时间：2018-04-30 02:16

本文选题：鹅出血性多瘤病毒 + 樱桃谷鸭　；参考：《中国畜牧兽医》2017年04期

【摘要】：为明确福建地区是否存在鹅出血性多瘤病毒(goose hemorrhagic polyomavirus,GHPV)感染,本研究对2016年福建省临床送检测19份鸭组织样品进行GHPV检测,结果从1例樱桃谷鸭中检测到GHPV感染阳性(记为GHPV-FJ201601株)。随后根据GenBank中GHPV参考株序列特征,设计针对GHPV的VP3基因特异性引物,利用PCR技术扩增获得GHPV的VP3基因片段。结果显示,GHPV-FJ201601株的VP3基因全长为654bp,编码217个氨基酸。对编码的VP3蛋白分析发现其理论等电点为9.37,带负电荷氨基酸为23个(Asp+Glu),带正电荷氨基酸为28个(Arg+Lys);不稳定指数为38.43,是一个稳定蛋白;脂肪系数64.33,总平均疏水性指数为-0.744;该蛋白无典型的信号肽切割位点;其亚细胞定位类型为核定位;对其进行磷酸化分析发现,该蛋白存在14个氨基酸磷酸化位点。核苷酸同源性分析显示,不同来源GHPV代表株VP3基因相互之间核苷酸同源性均较高,均不低于99.7%。VP3基因遗传进化结果显示,GHPV-FJ201601株和鹅源GHPV分离株(匈牙利14234株与法国Toulouse Goose 2000株)处于同一遗传进化分支,与鸭源分离株(法国Toulouse Muscovy Duck 2008株、法国Toulouse Mule Duck 2008和中国106株)却处于不同的遗传进化分支,但所有GHPV分离株VP3基因遗传进化均较近。本研究首次证实福建地区樱桃谷鸭群中存在GHPV感染,为丰富不同地区与宿主的GHPV分子流行病学数据提供参考。
[Abstract]:In order to find out whether goose hemorrhagic polytumour virus (Goose hemorrhagic polyomavirus) infection exists in Fujian province, 19 duck tissue samples from Fujian Province in 2016 were tested for GHPV. The results showed that GHPV infection was positive in one Cherry Valley duck (known as GHPV-FJ201601 strain). Then according to the sequence characteristics of GHPV reference strain in GenBank, the specific primers for VP3 gene of GHPV were designed, and the VP3 gene fragment of GHPV was amplified by PCR technique. The results showed that the VP3 gene of GHPV-FJ201601 strain was 654 BP, encoding 217 amino acids. The analysis of the encoded VP3 protein showed that the theoretical isoelectric point was 9.37, the negatively charged amino acid was 23 Amino acids and the positively charged Amino Acid was 28 Arg LysA, and the instability index was 38.43, which was a stable protein. The fat coefficient was 64.33, the total average hydrophobic index was -0.744; the protein had no typical signal peptide cleavage site; its subcellular localization type was nuclear localization; the phosphorylation analysis showed that there were 14 amino acid phosphorylation sites in the protein. Nucleotide homology analysis showed that the nucleotide homology of VP3 gene from different GHPV representative strains was high. The genetic evolution of GHPV-FJ201601 was not lower than that of 99.7%.VP3 gene. The results showed that 14234 strains of GHPV isolated from geese (14234 strains from Hungary and 2000 strains of Toulouse Goose from France) were in the same phylogenetic branch as duck isolates (2008 strains of Toulouse Muscovy Duck in France). In France, Toulouse Mule Duck 2008 and China 2008 were in different branches of genetic evolution, but all GHPV isolates had close genetic evolution of VP3 gene. This study first confirmed the existence of GHPV infection in Cherry Valley Duck population in Fujian Province, which provides a reference for enriching the molecular epidemiological data of GHPV in different regions and hosts.
【作者单位】：福建省农业科学院畜牧兽医研究所福建省畜禽疫病防治工程技术研究中心福建省禽病防治重点实验室;
【基金】：国家水禽产业技术体系(CARS-43) 国家自然科学基金(31602068) 福建省畜禽疫病防控技术重大研发平台(2014N2003) 福建省农业科学院畜牧兽医研究所基金(MYQJ2015(S)-3) 福建省属公益类项目(2015R1023-7) 福建省自然科学基金(2015J01114)
【分类号】：S852.65

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