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[浙江省鸽圆环病毒全基因组的克隆与分析]

[Cloning and analysis of the complete genomes of pigeon circovirus from Zhejiang Province].

作者信息

Yu Xu-Ping, Zhu Chun, Zheng Xin-Tian, Mu An-Xiong, Yu Hong-Tao

机构信息

College of Animal Sciences, Zhejiang University, Hangzhou 310029, China.

出版信息

Bing Du Xue Bao. 2009 Sep;25(5):355-61.

Abstract

Pigeon circovrius (PiCV) is a member of circovirus, which is usually regarded as an immunosuppression agent. There were reports that pigeons infected by PiCV showed symptoms of lethargy, weight loss, vomiting, diarrhea, respiratory distress, etc. In this study, we established a PCR method for the detection of PiCV DNA. Samples from 5 different farms in Zhejiang Province were examined and samples from a farm in Hangzhou were positive. Furthermore, the genomic segments of 2 strains of PiCV were amplified, cloned and sequenced using designed primers and the complete genomes of the strains were then assembled and named as PiCV-zj1 and PiCV-zj2, respectively. The sequences were deposited in GenBank under the GenBank Accession number of DQ090945 and DQ090944, respectively. Sequence Analysis had shown that the complete genomes of 2 strains of PiCV from Zhejiang Province had 2 039 nucleotides totally in length and common characters of circovirus such as a stem-loop structure and conserved motifs for Rep protein, which were supposed to be related to the replication of the virus. Pairwise comparisons showed that the nucleotide sequence of the genome of PiCV strains from Zhejiang Province had 86%-89.1% identities to that of 11 published PiCV strains, and that the amino acid identities of the replication-associated protein (Rep) and capsid protein (Cap) displayed 92.1%-94.7% and 76.6%-81.4%, respectively. A phylogenic tree was built using PHYLIP with bootstrap support for 1 000 replicates. The result showed that 10 strains from Europe and America formed one big branch and the others from Zhejiang Province and Australia formed the other two, respectively. This was the first report on the detection and full genome sequencing of PiCV in China.

摘要

鸽圆环病毒(PiCV)是圆环病毒属的成员,通常被视为一种免疫抑制剂。有报道称,感染PiCV的鸽子表现出嗜睡、体重减轻、呕吐、腹泻、呼吸窘迫等症状。在本研究中,我们建立了一种检测PiCV DNA的PCR方法。对浙江省5个不同养殖场的样本进行了检测,其中来自杭州一个养殖场的样本呈阳性。此外,使用设计的引物扩增、克隆并测序了2株PiCV的基因组片段,然后组装了这两个毒株的完整基因组,分别命名为PiCV-zj1和PiCV-zj2。这些序列分别以GenBank登录号DQ090945和DQ090944保存在GenBank中。序列分析表明,来自浙江省的2株PiCV的完整基因组全长共2039个核苷酸,具有圆环病毒的共同特征,如茎环结构和Rep蛋白的保守基序,这些特征被认为与病毒的复制有关。成对比较显示,来自浙江省的PiCV毒株基因组核苷酸序列与11株已发表的PiCV毒株的核苷酸序列同一性为86%-89.1%,复制相关蛋白(Rep)和衣壳蛋白(Cap)的氨基酸同一性分别为92.1%-94.7%和76.6%-81.4%。使用PHYLIP构建了一个有1000次重复自展支持的系统发育树。结果表明,来自欧美的10个毒株形成一个大分支,来自浙江省和澳大利亚的其他毒株分别形成另外两个分支。这是中国首次关于PiCV检测和全基因组测序的报道。

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