3株鸡传染性贫血病毒的全基因组序列分析

Complete Genomic Sequences Analysis of three chicken infection anemia viruses

Key words:  Chicken infection anemia virus  complete gene  VP1  genetic evolution

摘要:

通过对3株鸡传染性贫血病毒（CIAV）的全基因组序列比较分析，部分表明广西南宁鸡群CIAV毒株的遗传变异特征。将阳性CIAV的组织样品进行DNA抽提后，运用PCR方法分段扩增CIAV的全基因组，将获得的PCR产物进行基因克隆并进行阳性克隆菌鉴定后送测序。将所获得的基因序列进行拼接成CIAV基因组全长后，应用LaserGene7.1和MEGA4.1软件对CIAV全基因、Viral protein 3（VP3）、Viral protein 1（VP1） 和Viral protein 2（VP2）基因序列进行核苷酸、氨基酸同源性分析和遗传进化分析；并对VP1、VP2和VP3蛋白上与毒力、蛋白磷酸酶活性和细胞凋亡相关的氨基酸位点进行分析。结果获得3株CIAV的全基因序列，分别命名为GX1801、GX1804和GX1810；CIAV全基因遗传进化分析表明GX1801、GX1804和GX1810均同属于Group A群，与国内强毒株GD-103和GD-104毒株的亲缘关系较近；基于VP1基因构建的遗传进化树与全基因构建的进化树最相似；GX1801、GX1804和GX1810 VP1蛋白上的与毒力相关位点第75、89、125、141、144、394位氨基酸位点与日本强毒株C368株、国内强毒株GD-103和GD-104株在同一位点保持一致；GX1801、GX1804和GX1810 VP2蛋白上与磷酸酶活性相关的基序（I94CNCGQFRKH103）均未发生变异；GX1801、GX1804和GX1810 VP3蛋白与诱导细胞凋亡相关的重要区域（VP3蛋白N端结构域的第1-69位氨基酸）均高度保守。本研究对3株CIAV全基因组进行测定并进行基因分析，获得了其基因组特征，为进一步研究广西CIAV的分子流行和致病性提供参考。

Abstract:

To investigate the characteristic of three complete genomic sequences of chicken infection anemia virus (CIAV) from Nanning, Guangxi province, the CIAV complete genomes from the positive samples were amplified by PCR after DNA extraction from liver samples of commercial broilers. LaserGene7.1 and MEGA4.1 softwares were used to analyze the nucleotide / amino acid homology and genetic evolution of CIAV complete genomes, Viral protein 3 (VP3), Viral protein 1 (VP1) and Viral protein 2 (VP2); And the amino acid sites related to viral virulence, protein phosphatase activity and apoptosis of VP1, VP2 and VP3 were analyzed. The results showed that three CIAV complete genome sequences were successfully obtained, named GX1801, GX1804 and GX1810, respectively; Genetic evolution analysis showed that GX1801, GX1804 and GX1810 belonged to the Group A and were closely related to the domestic virulent strain GD-103 and GD-104; The phylogenetic trees results showed that the phylogenetic tree based on VP1 gene was the most similar to that based on the complete genome; the amino acid sites at 75aa, 89aa, 125aa, 141aa, 144aa and 394aa of VP1 protein of GX1801, GX1804 and GX1810 were the same as those of C368, GD-103 and GD-104 strain; There was no variation of the phosphatase activity related motifs (I94CNCGQFRKH103) on VP2 protein in GX1801, GX1804 and GX1810, and the important regions (the amino acid sites at 1 to 69 of N-terminal domain) related to apoptosis induction of VP3 protein in GX1801, GX1804 and GX1810 VP3 were highly conserved. In this study, three complete genomes of CIAV obtained from Nanning city of Guangxi were analyzed, and the CIAVs were speculated to maybe highly pathogenic, which provide reference for the molecular epidemiological investigation of CIAV in Guangxi.