Cloning and Characterization of the Polymerase Gene of Porcine Coronavirus

LI Jian-qiang; LIU Ji-xing; CHENG Jie; LAN Xi; HU Yong-hao; WU Run

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December 2006

LI Jian-qiang, LIU Ji-xing, CHENG Jie, LAN Xi, HU Yong-hao and WU Run. Cloning and Characterization of the Polymerase Gene of Porcine Coronavirus[J]. Virologica Sinica, 2006, 21(6): 551-555.

Citation: LI Jian-qiang, LIU Ji-xing, CHENG Jie, LAN Xi, HU Yong-hao, WU Run. Cloning and Characterization of the Polymerase Gene of Porcine Coronavirus .VIROLOGICA SINICA, 2006, 21(6) : 551-555.

猪源冠状病毒聚合酶基因的克隆及特性分析

李建强 ^1,2 ,
柳纪省 ^1*,, ,
胡永浩 ,

1.
1.中国农业科学院兰州兽医研究所，家畜疫病病原生物学国家重点实验室，农业部畜禽病毒学重点开放实验室，甘肃兰州 730046
2.
甘肃农业大学动物医学院，甘肃兰州 730070

通讯作者： 柳纪省, ;

摘要

参照GenBank中Purdue株序列对TGEV TS株聚合酶基因（ORF1）设计特异性引物,经RT-PCR扩增获得了20054bp的片段，与预期大小相符。TS株与Pur46-MAD株的ORF1核苷酸和氨基酸序列同源性分别为98.8%和99.0%,与FIPV、PEDV、HCV299E及SARS氨基酸同源性分别为88%、57%、57%和45%。不同冠状病毒比较结果显示RdRp有很高的保守性而且有重要的作用，序列分析标明TS株在ORF1a和ORF1b重叠区有核糖体剪切位点UUUAAAC，且相应区域RNA二级结构形成3个茎环结构。
- 猪源冠状病毒
- , 聚合酶基因
- , 克隆
- , 特征分析

Cloning and Characterization of the Polymerase Gene of Porcine Coronavirus

LI Jian-qiang ^1,2 ,
LIU Ji-xing ^1*,, ,
CHENG Jie ,
LAN Xi ¹ ,
HU Yong-hao ² ,
WU Run ²

1.
1. Key laboratory of Animal Virology of Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China
2.
College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China

Corresponding author: LIU Ji-xing,

Abstract

The polymerase gene, a non-structural gene from strain TS of transmissible gastroenteritis virus (TGEV), was amplified by RT-PCR primers designed based on the Purdue nucleotide sequence in GenBank. The expected 20054 bp product was obtained. The nucleotide sequence of ORF1 of TS shared nucleotide and amino acid identities of 98.8% and 99.0%, respectively with that of strain Pur46-MAD. The identity at the amino acid level for the ORF1 between TS and FIPV, PEDV, HCV299E,SARS was 87%、57%、57%、45%, respectively. RdRp was regarded to have an important role in replication and the results indicated that it was a conserved protein. The data also showed that there was a ribosomal slippage site UUUAAAC and three stem-loop structures in the ORF1a and ORF1b overlapping regions.
- Porcine Coronavirus
- , Polymerase gene
- , Cloning
- , Characterization analysis

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Cloning and Characterization of the Polymerase Gene of Porcine Coronavirus

Corresponding author: LIU Ji-xing,

1. 1. Key laboratory of Animal Virology of Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China
2. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China

Abstract: The polymerase gene, a non-structural gene from strain TS of transmissible gastroenteritis virus (TGEV), was amplified by RT-PCR primers designed based on the Purdue nucleotide sequence in GenBank. The expected 20054 bp product was obtained. The nucleotide sequence of ORF1 of TS shared nucleotide and amino acid identities of 98.8% and 99.0%, respectively with that of strain Pur46-MAD. The identity at the amino acid level for the ORF1 between TS and FIPV, PEDV, HCV299E,SARS was 87%、57%、57%、45%, respectively. RdRp was regarded to have an important role in replication and the results indicated that it was a conserved protein. The data also showed that there was a ribosomal slippage site UUUAAAC and three stem-loop structures in the ORF1a and ORF1b overlapping regions.