首页|期刊导航|中国农业科学(英文版)|Cloning and Homology Comparison of S Gene for Isolate TH-98 of Porcine Transmissible Gastroenteritis Virus
中国农业科学(英文版)2003，Vol.2Issue(3)：314-320,7.
Cloning and Homology Comparison of S Gene for Isolate TH-98 of Porcine Transmissible Gastroenteritis Virus
Cloning and Homology Comparison of S Gene for Isolate TH-98 of Porcine Transmissible Gastroenteritis Virus
摘要
Abstract
TH-98 isolate of transmissible gastroenteritis virus (TGEV) was propagated and harvested onswine testicle (ST) monolayer ceil. Two pairs of primers were designed to amplify S gene by RT-PCR accord-ing to the published sequence of TGEV'S gene cDNA with Oligo version 4.1 and DNasis software. The productsof PCR were named Sa and Sb, of 2.3 kb and 2.1 kb respectively. Sa was inserted in EcoR I and Kpn I sitesafter Sb was cloned in Kpn I and Pst I multiple cloning sites of the same pUC18 plasmid. The recombinantpUC-S plasmid was identified and analyzed by corresponding restriction endonuclease and nested PCR on thebasis of the genetic sites of S gene and pUC18 plasmid, which was identified as S gene of TGEV. RecombinantpUC-S was sequenced and analyzed in comparison with the other strains. Gene sequence comparison indicatedthat TH-98 shared 99, 97, 98, 97 and 94% identities with Purdue-115(US), Miller(US), TO14(Japan),FS772(British), 96-1933(British), respectively, their deduced amino acid homology was 99, 97, 97, 96 and93% correspondingly. In addition, the analysis report verified that pUC-S owned a complete open readingframe (ORF) including initiation codon, signal sequences, remaining sequences and termination codon aswell. Therefore, the results affirmed that S gene of TGEV TH-98 was extremely conservative.关键词
Transmissible gastroenteritis virus/S gene/Cloning/Homology ComparisonKey words
Transmissible gastroenteritis virus/S gene/Cloning/Homology Comparison分类
农业科技引用本文复制引用
REN Xiao-feng,LI Yi-jing,LIU Bao-quan..Cloning and Homology Comparison of S Gene for Isolate TH-98 of Porcine Transmissible Gastroenteritis Virus[J].中国农业科学(英文版),2003,2(3):314-320,7.基金项目
We are grateful to all those who supplied withexperimental materials in this study. Viruses wereisolated with the cooperation of other colleagues ofPreventive Veterinary College of Northeast Agricultural University, China. ST cells were kindlyprovided by （）
