【作者】 希尼尼根； 程安春； 汪铭书； 陈孝跃； 文明； 周伟光； 郭宇飞； 陈斌； 贾仁勇；

【Author】 XininiGeng, Cheng Anchun, Wang Mingshu ,et al (Sichuan Agricultural University, Yaan, Sichuan, 625014)

【机构】 四川农业大学动物科技学院动物疫病和食品安全评价四川省重点实验室；

【摘要】 本研究根据genbank中猪繁殖与呼吸综合征病毒(PRRSV)VR-2332株的ORF5基因序列设计并合成了1对引物,对2株PRRSV四川分离株(SC1和SC2)进行RT-PCR扩增出了1条包括完整ORF5基因的739bp的cDNA片段。将该扩增片段经T-A连接插入pGEM-Teasy载体中,用重组质粒转化大肠杆菌DH5a,获得重组质粒转化菌。对重组质粒进行酶切鉴定,证明插入的cDNA片段与RT-PCR扩增的cDNA片段大小相符。同时对此cDNA片段进行全序列测定,并与美洲代表株VR-2332株及欧洲代表株LV做了比较。结果表明,2株四川分离株ORF5基因序列与VR-2332株的十分相似,其核苷酸同源性达到98-99%,氨基酸同源性为99%,而与LV株,则相差甚远,其核苷酸及氨基酸同源性分别为57%和54%。本研究为四川地区PRRS的诊断和防制提供了依据。更多还原

【Abstract】 Porcine reproductive and respiratory syndrome (PRRS) is a newly recognized pig disease. The causative agent of PRRS is porcine reproductive and respiratory syndrome virus (PRRSV). In this study a pair of primers was designed according to the VR-2332 strain and a fragment of two different SiChuan isolates amplified respectively by RT-PCR and cloned into T-easy vector, then two recombinants respectively harboring the ORF5 gene of the two SjChuan isolates was designated as T-SC1-ORF5 and T-SC2-ORF5. The purified recombinant was subjected to sequencing. The ORF5 gene of both SiChuan isolates displayed 98-99% nucleotide identities and 99% amino acid identities with strain ATCC VR-2332, and 57% nucleotide identities and 54% amino acid identities with strain LV. The hydrophobic regions are nearly identical between two SiChuan isolates and RV 2332 strain. The results indicate that the rwo SiChuan isolates are closely related to VR-2332 strain, but not so to LV strain. As ORF5 gene of PRRSV is relatively conservative, the cloned ORF5 gene in the study provided a scientific basis for diagnostic use, as well as possible vaccine candidate.更多还原