猪圆环病毒2型2d基因型毒株的全基因组序列分析

发布时间：2018-02-21 05:24

本文关键词： 猪圆环病毒型分离鉴定全基因序列 PCVd基因型　出处：《中国兽医科学》2017年12期 　论文类型：期刊论文

【摘要】：根据Gen Bank中猪圆环病毒2型(PCV 2)基因序列,设计特异性引物,用PCR方法从福州郊区某猪场疑似PMWS仔猪病料处理后接种育传三代的PK-15细胞中扩增1个分离毒株的全基因组,并将其克隆到p MD-18T载体上,对鉴定为阳性的重组质粒送检测序。对该分离株的基因全序列及其开放阅读框2(ORF2)进行同源性分析及遗传进化关系分析。PCR鉴定结果显示,本试验中分离到的病毒属于PCV2,命名为PCV2-FZ株。全基因组序列分析显示,分离株全基因组大小为1 767 bp,含有11个开放性阅读框,其中ORF2大小为705 bp,共编码234个氨基酸。PCV2-FZ株与参考毒株基因全序列的核苷酸同源性为94.3%~99.4%,与SD毒株同源性最高,为99.4%;与DK1980PMWSfree和DK1990PMWSfree毒株同源性最低,为94.3%。分离株与参考毒株ORF2核苷酸同源性为89.2%~99.4%,其中与SD毒株同源性最高,为99.4%,与DK1980PMWSfree毒株和DK1987PMWSfree毒株同源性最低,均为89.2%。全基因组与ORF2序列遗传进化树分析均表明,PCV2-FZ株属于PCV2d基因型。本研究从福建省首次分离并鉴定了1株PCV2d型毒株,这对PCV2流行病学研究和遗传变异分析具有重要意义。
[Abstract]:According to the sequence of porcine circovirus type 2 (PCV2) gene in Gen Bank, a specific primer was designed and the whole genome of a isolated virus strain was amplified by PCR from PK-15 cells of suspected PMWS piglets in a pig farm in the suburb of Fuzhou. It was cloned into p MD-18T vector, and the recombinant plasmid identified as positive was submitted for sequencing. Homology analysis and genetic evolution analysis showed that the whole gene sequence of the isolate and its open reading box 2 ORF2) were analyzed. The whole genome sequence analysis showed that the whole genome size of the isolate was 1 767 BP, containing 11 open reading frames. The nucleotide homology of ORF2 was 705 BP, encoding 234 amino acids. PCV2-FZ and reference strain. The nucleotide homology was 94.33.99. 4, the highest homology with SD strain was 99.4, and the homology with DK1980PMWSfree and DK1990PMWSfree strains was the lowest. The nucleotide homology of ORF2 between the isolated strain and reference strain was 89.2and 99.4, the highest homology with SD strain was 99.4, and the homology with DK1980PMWSfree strain and DK1987PMWSfree strain was the lowest. The whole genome and ORF2 sequence genetic phylogenetic tree analysis showed that PCV2-FZ strain belonged to PCV2d genotype. One PCV2d strain was isolated and identified from Fujian Province for the first time, which is of great significance for the epidemiological study of PCV2 and the analysis of genetic variation.
【作者单位】：福建农林大学动物科学学院;福建省兽医中药与动物保健重点实验室;
【基金】：国家自然科学基金青年基金项目(31502110):猴头菇多糖对氧化应激状态下猪小肠上皮通透性的调节及其机制
【分类号】：S852.651

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