四川农业大学学报 ›› 2007, Vol. 25 ›› Issue (02): 127-130.doi: 10.16036/j.issn.1000-2650.2007.02.004

• 研究论文 • 上一篇    下一篇

猪圆环病毒1型和2型四川分离株全基因克隆分析

朱玲, 郭万柱, 徐志文, 陈杨, 周远成   

  1. 四川农业大学 动物科技学院, 四川 雅安 625014
  • 收稿日期:2007-04-17 出版日期:2007-06-30 发布日期:2017-03-03
  • 基金资助:
    四川省科技厅生物技术专项;四川省教育厅重点专项(2004D004)。

Cloning and Sequencing of Complete Genome of Porcine Circovirus Type 1 (PCV1)and Porcine Circovirus Type 2 (PCV2)

ZHU Ling, GUO Wan-zhu, XU Zhi-wen, CHEN Yang, ZHOU Yuan-cheng   

  1. College of Animal and Technology, Sichuan Agricultural University, Yaan 625014, Sichuan, China
  • Received:2007-04-17 Online:2007-06-30 Published:2017-03-03

摘要: 参照Genbank收录猪圆环病毒(porcine circovirus,PCV)1、2型序列,设计两对特异性引物,分别扩增猪圆环病毒1型和2型四川分离株(PCV1-YA1株和PCV2-SC株)全基因组,获得了预期目标大小一致扩增产物,分别将其克隆于PMD18-T载体,分别命名为PMD18-T-PCV1-YA与PMD18-T-PCV2-SC。经酶切鉴定和序列测定证实,成功克隆了PCV1-YA1株1759bp和PCV2-SC株1767 bp的全基因组序列。所得序列与GenBank中的国内外其它PCV1和PCV2毒株进行比较分析,其同源性分别达到98.8%~99.9%与93.6%~98.6%;而PCV1-YA1株和PCV2-SC株之间的同源性则仅为69.2%,进一步分析两个毒株的主要阅读框架,毒株间ORF1核苷酸序列及推导的氨基酸同源性均为85.3%,而ORF2的核苷酸序列及推导的氨基酸同源性分别为66%和65.2%。推导显示两毒株之间ORF1编码产物疏水性区域分布有相似之处,而ORF2编码产物的跨膜区存在差异。

关键词: 猪圆环病毒, 全基因, 克隆, 序列分析

Abstract: One pair of primers was designed based on the Genbank published sequence of Porcine Circovirustype 1(PCV1) to amplify the complete PCV1 genome of YA1 strain, resulting a 1759 bp PCR product.Then cloned the PCR product into pMD18-T vector to obtain the recombinant vector PMD18-T-PCV1-YA.Enzyme and sequencing analysis shows that the complete sequence of PCV1 was successfully cloned.Sequenced similarity analysis show ed that PCV1-YA1 strain shared 98.8%~99.9% nucleotide sequence identity with the other six strains in GenBank.Complete PCV2(1767 bp) sequence of SC strain was cloned into PMD18-T-PCV2-SC successfully using the same method, and sequenced similarity analysis showed that PCV2-SC strain shared 93.6%~98.6% nucleo tide sequence identity with other strains in GenBank.The nucleotide sequence identity between PCV1-YA1 and PCV2-SC is only 69.2%.The two major open-reading frame analysis shows that the nucleotide sequence identity of ORF1 and ORF2 between PCV1-YA1 and PCV2-SC are 85.3% and 66%, and their deduction amino acids identity are 85.3% and 65.2% respectively.The hydrophobic prediction revealed that PCV1 ORF1 is similar to PCV2 ORF1, and the transmembrane prediction revealed ORF2 are difference between the two strains.

Key words: porcine circovirus, com plete genome, clone, sequenced similarity analysis

中图分类号: 

  • S858.280.43