›› 2015, Vol. 42 ›› Issue (7): 1668-1673.doi: 10.16431/j.cnki.1671-7236.2015.07.007

• 生物技术 • 上一篇    下一篇

猪细环病毒1a型ORF1基因的克隆与序列分析

陈鹏强, 胡崇伟, 林群群, 陈秋勇, 修金生   

  1. 福建农林大学动物科学学院, 福州 350002
  • 收稿日期:2015-01-05 出版日期:2015-07-20 发布日期:2015-07-23
  • 通讯作者: 修金生 E-mail:xiujinsheng@163.com
  • 作者简介:陈鹏强(1991-),男,福建南安人,硕士生,研究方向:猪病的防控,E-mail:chenpq1991@163.com
  • 基金资助:
    福建省农业科学技术项目(2014-01);福建省自然科学基金项目(2013J05042)

Cloning and Sequence Analysis of ORF1 Gene of Porcine Torque Teno sus Virus Type 1a

CHEN Peng-qiang, HU Chong-wei, LIN Qun-qun, CHEN Qiu-yong, XIU Jin-sheng   

  1. College of Animal Science, Fujian Agriculture and Forestry University, Fuzhou 350002, China
  • Received:2015-01-05 Online:2015-07-20 Published:2015-07-23

摘要: 为明确猪细环病毒(porcine Torque teno sus virus,PTTSuV)1a型福建株ORF1基因的特征,本研究采用分段扩增的办法从PTTSuV 1a型感染阳性粪便中扩增PTTSuV 1a型福建株ORF1基因,对PCR扩增产物胶回收克隆测序后利用SeqMan软件拼接出完整的PTTSuV 1a型福建株ORF1基因,通过分子生物学软件对其编码蛋白进行生物信息学分析。结果表明,PTTSuV 1a型福建株ORF1基因全长为1 947 bp,编码648个氨基酸,其理论等电点为10.06。核苷酸同源性比对结果表明,本试验PTTSuV 1a型福建株ORF1基因和PTTSuV 1a型TTV1 Bj2-1株(GenBank登录号:HM633243)同源性最高,达99.7%。利用Mega 6.06绘制其遗传进化树,可见PTTSuV 1a型ORF1基因在遗传进化上呈两个大的遗传进化分支(分支Ⅰ和分支Ⅱ),分支Ⅰ又可分为两个小的分支(Ⅰa和Ⅰb),本研究为丰富福建源PTTSuV 1a型分子流行病学数据库奠定基础。

关键词: 猪细环病毒1a型; ORF1基因; 克隆; 序列分析

Abstract: In order to know the ORF1 gene of the Fujian isolates of porcine Torque teno sus virus (PTTSuV) type 1a,the study amplified the ORF1 gene of PTTSuV type 1a from the DNA isolated in the feces with diarrhea in Fujian province.The target PCR fragments were cloned and sequenced,and spliced with SeqMan software.The results showed that the cloned ORF1 gene of PTTSuV type 1a was 1 947 bp in length,coding an open reading frame (ORF) with 648 amino acids,its theoretical isoelectric point was 10.06.The nucleotide sequence and the amino acid sequence deduced from the gene were analyzed by the bioinformatics software.Compared with the PTTSuV type 1a ORF1 gene from GenBank,the sequenced gene shared the highest homology with TTV1 Bj2-1 strain (GenBank accession number:HM633243) at 99.7%.The phylogenetic tree was constructed with Mega 6.06,the PTTSuV type 1a could be divided into 2 different phylogenetic clusters (clusterⅠ and cluster Ⅱ),the cluster Ⅰ also had two distinguish branch (Ⅰa and Ⅰb).The results of this study could enrich the genomic characterization of PTTSuV type 1a in Fujian.

Key words: porcine Torque teno sus virus type 1a; ORF1 gene; clone; sequence analysis

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