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两株不同来源的猪圆环病毒1型ORF2基因的
克隆与序列分析
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张福良1，2，孙丽华1，3，宋长绪1，杨鸣琦2，郝永清3，刘燕玲1，武占银4
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（1.广东省农业科学院 兽医研究所，广东 广州510640; 2.西北农林科技大学 动物科技学院，
陕西 杨凌712100； 3.内蒙古农业大学 动物科学与医学学院，内蒙古 呼和浩特010018；
4.宁夏动物防疫站，宁夏 银川750000）
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摘要：
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根据GenBank中猪圆环病毒1型（PCV1）ORF2基因序列，设计了1对引物，应用PCR方法从PK-15细胞和疑似患断奶仔猪多系统衰竭综合征（PMWS）的仔猪组织病料中分别扩增出了PCV1 ORF2全基因（702bp）。将此基因片段克隆入pMD 18-T载体，筛选获得了重组质粒pMD-PCV1-ORF2，并对其进行了测序与分析。结果表明：所克隆的PK-15来源和组织病料来源的ORF2基因的同源性为99.4%,二者与GenBank中登录的PCV1 ORF2基因的同源性为973%～99.9%,与PCV2 ORF2基因的同源性为67.9%～68.4%,二者编码的蛋白在几个功能区比较保守。抗原表位预测表明，PCV1 ORF2编码的蛋白具有良好的免疫原性。 |
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关键词：
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猪圆环病毒1型； ORF2基因； 克隆； 序列分析 |
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中图分类号：S 852.659.2 文献标识码：
A 文章编号：1673-4696(2006)04-0282-05 |
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Cloning and sequencing analysis of ORF2 gene of
porcine circovirus type 1 |
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ZHANG Fu-liang1,2，SUN Li-hua1,3，SONG
Chang-xu1，YANG Ming-qi2，
HAO Yong-qing3 , LIU Yan-ling1, WU Zhan-yin4 |
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(1. Veterinary Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou 510640,China;
2.Northwest Sci-Tech University of Agriculture and Forestry, Yangling 712100,China; 3.College of Animal
Science and Medicine, Nei Monggol Agricultural University,Hohhot 010018,China;
4.Ningxia Animal Epidemic Prevention Station, Yinchuan 750000, China ) |
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Abstract: The ORF2 gene was amplified from PK-15 cells and tissues of piglets suffered from PMWS,respectively using the primers designed according to the OFR2 sequences published in GenBank.The resultant PCR products were cloned into pMD18-T vectors and identified by restriction digestion using endonucleases XhoⅠ and KpnⅠ.Nucleotide sequences of the PCV1 ORF2 genes were compared with that of other PCV1 ORF2 and PCV2 ORF2 genes. The results indicated that nucleotide homology between ORF2 genes cloned from PK-15 cells and pig tissues was 99.4%,nucleotide homologies between the cloned PCV ORF2 genes and the published PCV1 ORF2 genes in GenBank were from 97.3% to 99.9%,and homologies between the cloned PCV ORF2 genes and the published PCV2 ORF2 genes in GenBank were from 67.9% to 68.4%.Several conserved functional domains were identified in the cloned PCV1 ORF2 genes. Predication of antigenicity showed that the protein translated by the PCV1 ORF2 genes may have good immunogenicity. |
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Key words: porcine circovirus type 1; ORF2 gene; cloning; sequence analysis |
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