猪繁殖与呼吸综合征病毒S1株基因组序列测定与分析.pdfVIP

第 21 卷 5 期 中 国 病 毒 学 21 （5 ）： 490- 493 2006 年 9 月 VIROLOGICA SINICA September 2006 * 猪繁殖与呼吸综合征病毒 S1 株基因组序列测定和分析 李玉峰，姜 平** ，许家荣，蒋文明 （南京农业大学动物医学院，江苏南京 210095 ） Genomic Sequencing of S1 Strain of a Porcine Reproductive and Respiratory Syndrome Virus Isolate ** LI Yu-feng ，JIANG Ping ，XU Jia-rong ，JIANG Wen-ming (College of Animal Medicine, Nanjing Agricultural University, Nanjing 210095, China) Abstract: Four gene fragments of S1 strain of porcine reproductive and respiratory syndrome virus were amplified by RT-PCR and cloned into the pCR-XL-TOPO vector for sequencing. Authentic 5′ and 3′ terminal fragments of S1 strain were obtained by 5′ -Full RACE Core Set and 3′ -Full RACE Core Set and cloned into the pMD-18T vector and sequenced. The complete nucleotide sequence of S1 strain genome was obtained by sequential splicing of each fragment. The results showed that the genome of S1 strain was 15441 bp in length, containing nine open reading frames (ORFs), with 189 nt in the 5’ UTR and 181 nt in the 3’ UTR, including 30 nt poly (A) tail. The genome of PRRSV S1 strain showed 99.5%, 99.6% and 90.8% of nucleotide identity with ATCC VR-2332, BJ-4 and CH-1a, respectively. Key words: PRRSV; Genome; Sequencing and analysis 摘要：应用 RT-PCR 方法分段扩增出 PRRSV 上海分离株 S1 毒株的 4 条基因大片段，扩增后的产物分别克隆于 pCR-XL-TOPO 载体鉴定后测序，同时应用 RACE 方法对 S1 毒株的 3′和 5′ 基因末端进行了成功的扩增并克隆 于 pMD-18T 载体进行测序，按顺序将这些序列进行拼接得到 PRRSV S1 株全基因组 cDNA 序列。测序结果表 明PRRSV S1 株基因组全长 15441 bp，包含 9 个开放式阅读框，5′UTR 含有