猪细环病毒1a型ORF1基因的克隆与序列分析  

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

作　　者：陈鹏强[1] 胡崇伟[1] 林群群[1] 陈秋勇[1] 修金生[1] 

机构地区：[1]福建农林大学动物科学学院,福州350002

出　　处：《中国畜牧兽医》2015年第7期1668-1673,共6页China Animal Husbandry & Veterinary Medicine

基　　金：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)

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 947bp,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TTV1Bj2-1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PTTSuV1a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。In order to know the ORF1 gene of the Fujian isolates of porcine Torque teno sus virus （PTTSuV） type la,the study amplified the ORF1 gene of PTTSuV type la from the DNA isola- ted in the feces with diarrhea in Fujian province. The target PCR fragments were cloned and se- quenced,and spliced with SeqMan software. The results showed that the cloned ORF1 gene of PTTSuV type la 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 se- quence deduced from the gene were analyzed by the bioinformatics software. Compared with the PTTSuV type la 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 la could be divided into 2 different phy- logenetic clusters （cluster I and cluster Ⅱ ） ,the cluster Ⅰ also had two distinguish branch （ I a and I b）. The results of this study could enrich the genomic characterization of PTTSuV type la in Fujian.

关 键 词：猪细环病毒1a型 ORF1基因 克隆 序列分析 

分 类 号：S852.65[农业科学—基础兽医学]