猪细环病毒1b型ORF3基因克隆及序列分析

中国畜牧兽医

猪细环病毒1b型ORF3基因克隆及序列分析

胡崇伟¹，陈如敬²，陈秋勇¹，林群群¹，陈鹏强¹，修金生¹

1.福建农林大学动物科学学院，福建福州 350002；2.福建省农业科学院畜牧兽医研究所，福建省畜禽疫病防治工程技术研究中心，福建福州 350013

修回日期:2014-04-04 出版日期:2014-08-20 发布日期:2014-08-22
通讯作者: 修金生（1957—），男，福建人，教授，硕士生导师，研究方向：猪病防治和生态养猪。E-mail：xiujinsheng@163.com
作者简介:修金生（1957—），男，福建人，教授，硕士生导师，研究方向：猪病防治和生态养猪。E-mail：xiujinsheng@163.com
基金资助:
福建省自然科学基金（2013J05042）。

Cloning and Sequence Analysis of ORF3 Gene of Porcine Torque Teno sus Virus 1b

HU Chong-wei¹, CHEN Ru-jing², CHEN Qiu-yong¹, LIN Qun-qun¹, CHEN Peng-qiang¹, XIU Jin-sheng¹

1. College of Animal Science, Fujian Agriculture and Forestry University, Fuzhou 350002, China; 2. Fujian Animal Diseases Control Technology Development Center, Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou 350013, China

Revised:2014-04-04 Online:2014-08-20 Published:2014-08-22

摘要/Abstract

摘要： 为明确福建省猪细环病毒（porcine torque teno sus virus，PTTSuV）1b型（PTTSuV-1b）ORF3基因的遗传进化特征，本研究根据GenBank中登录的PTTSuV-1b基因组特征设计特异性引物，对福建省某猪场患有仔猪断奶后多系统衰竭综合征(PMWS)的猪血清进行PTTSuV-1b分段扩增，并分别对PCR扩增产物进行胶回收后克隆测序，将测序结果经BLAST分析后进行序列拼接。试验结果表明，所扩增的目的片段编码有完整的PTTSuV-1b ORF3蛋白，全长为600 bp，编码有199个氨基酸。将获得的PTTSuV-1b型福建株与GenBank中PTTSuV-1b型的ORF3基因进行比对分析，其与FJ/China/2010/TTV2/2株核苷酸同源性最高，为99.7%，与西班牙PTTSuV-1b分离株TTV2_G43核苷酸同源性为97.3%，与SC株核苷酸同源性稍低，但也达94.0%；而与猪细环病毒K2型德国家猪分离株472142株核苷酸同源性仅为60.7%，与猪细环病毒1a型西班牙分离株PTTV1_1914株核苷酸同源性仅为46.8%。从遗传进化关系上看，PTTSuV-1b ORF3基因在遗传进化上呈2个大的遗传进化分支（分支Ⅰ和分支Ⅱ），本研究分离株处于分支Ⅰ。

关键词: 猪细环病毒1b型; ORF3基因; 序列分析

Abstract: In order to enrich the ORF3 target protein information of porcine torque teno sus virus 1b (PTTSuV-1b), two parts of ORF3 fragments were amplified by PCR method from sera of the piglets association with post-weaning multisystem wasting syndrome in Fujian province. Then the target PCR products were cloned and sequenced. The results demonstrated that PTTSuV-1b ORF3 gene was 600 bp in length, coding an open reading frame (ORF) with 199 amino acids. The nucleotide sequence and amino acid sequence deduced from the gene were analyzed by the bioinformatics software. Compared with the PTTSuV-1b ORF3 genes download from GenBank, the sequenced gene shared the highest homogeneity with FJ/China/2010/TTV2/2 strain at 99.7%, with the SC strain at 94.0%, with the Spain PTTSuV-1b TTV2_G43 isolates at 97.3%. However, the PTTSuV-1b ORF3 genes shared with the torque teno sus virus K2 472142 stain isolated from Germany at 60.7%, with the Spain PTTSuV-1a PTTV1_1914 isolates at 46.8%. According to the results of the phylogenetic relationship constructed with Mega, PTTSuV-1b had two distinguish branches (branchⅠ and branch Ⅱ), the Fujian isolates were at the branchⅠ.

Key words: porcine torque teno sus virus 1b; ORF3 gene; sequence analysis

胡崇伟，陈如敬，陈秋勇，林群群，陈鹏强，修金生. 猪细环病毒1b型ORF3基因克隆及序列分析[J]. 中国畜牧兽医.

HU Chong-wei, CHEN Ru-jing, CHEN Qiu-yong, LIN Qun-qun, CHEN Peng-qiang, XIU Jin-sheng. Cloning and Sequence Analysis of ORF3 Gene of Porcine Torque Teno sus Virus 1b[J]. .