猪繁殖和呼吸综合征病毒ORF7基因的克隆与序列分析  

Amplification and Sequence Analysis of ORF7 Gene of Porcine Reproductive and Respiratory Syndrome Virus

作　　者：吴国平[1] 刘冰心[2] 郭川[3] 曹敏杰[2] 苏文金[2] 

机构地区：[1]集美大学水产学院水产生物技术研究所,福建厦门361021 [2]集美大学生物工程学院,福建厦门361021 [3]厦门大学生命科学学院,福建厦门361005

出　　处：《集美大学学报（自然科学版）》2006年第3期217-222,共6页Journal of Jimei University：Natural Science

基　　金：福建省科技攻关计划重点项目(2003N083);厦门市科技攻关计划重点项目(3502Z20031054)

摘　　要：根据猪繁殖和呼吸综合征病毒(PRRSV)已发表的核苷酸序列,设计了1对特异性引物P1/P2,用RT-PCR扩增了PRRSV福建毒株FJ-1和FJ-2的全长核衣壳蛋白基因(ORF7),将扩增产物连接到pUCm-T载体并转化大肠杆菌DH10B,提取阳性重组质粒进行序列测定与分析.结果表明:FJ-1和FJ-2毒株ORF7基因均为372 bp,编码123个氨基酸,与美洲型参考毒株VR-2332及欧洲型参考毒株LV的核苷酸同源性分别为95.16%,94.35%和61.40%,60.65%,其推导的氨基酸同源性分别为95.93%,95.12%和56.49%,55.73%.研究表明,福建流行的FJ-1和FJ-2属于美洲型PRRSV毒株.Porcine Reproductive and Respiratory Syndrome Virus （PRRSV） strains FJ-1 and FJ-2 were isolated from a pig farm in Fujian province of China during an outbreak of PRRS. The full-length of ORF7 gene encoding the nucleocapsid protein （N） protein of FJ-1 and FJ-2 was amplified by RT-PCR with a pair of primers （P1/P2） and cloned into pUCm-T vector, and transformed to E. coli DH10B competent cell. The purified recombinants were subjected to sequencing and analysis. The ORF7 genes of FJ-1 and FJ-2 were both 372 bp and encoding 123 amino acids. They displayed 95. 16 %, 94. 35 % and 61.40 %, 60. 65 % nucleotide identities to strains VR-2332 （a reference strain of America） and LV （European genotype）, respectively. The deduced amino acids of FJ-1 and FJ-2 showed 95.93 %, 95.12 % and 56.49 %, 55.73 % identifies to strains VR-2332 and LV. The results shown here strongly suggested that PRRSV strains FJ-1 and FJ-2 belong to American genotype.

关 键 词：PRRSV 福建毒株 ORF7 序列分析 

分 类 号：S852.65[农业科学—基础兽医学] Q78[农业科学—兽医学]