【摘要】 山东某些养殖场发生以母猪流产或产死胎、木乃伊等为特症的流行性繁殖障碍病,怀疑为猪繁殖与呼吸综合症病毒(PRRSV)所致。自行设计了一对针对PRRSV的N基因的特异性引物P1和P2,通过RT-PCR技术对分别从潍坊和济南收集到的可疑病料进行检测,结果为阳性,并得到1条366bp的DNA片段。纯化此扩增产物,然后与PMD-18T载体连接,转化大肠杆菌DH5α。结果得到了1个ORF7基因与PMD-18T载体的重组质粒PMD-18T- ORF7。通过EcoRI/BamHI双酶切及PCR证明此重组质粒即为ORF7基因与PMD-18T载体的重组质粒,从而为ORF7基因及其表达的核表壳蛋白进一步研究奠定了基础。更多还原

【Abstract】 Recently,an infectious disease occured frequently in some pig farms in Shandong province. The characteristics of this epedemic are abortion,death of embryo and mummified fetuses,which is con sid- ered as the result of porcine reproductive and respiratory syndrome virus(PRRSV).With a specific N gene primers of PRRSV,it was identified that the samples collected from these farms in Weifang and Jinan were positive by RT-PCR based on it,so two primes P1 and P2 were designed.The fragments had been success- fully amplified from RNA of PRRSV from samples.The length of fragments is 366 base pair.The ampli- fied product was purified and ligated into PMD-18T vector,and finally transferred into E.coli DH5α,a re- combinant plasimid of ORF7 gene and PMD-18T vector were obtained.By the methods of digestion with EcoRI/BamHI and PCR,the recombinant plasimid was identified that it is recombined with ORF7 gene and PMD-18T vector.This research will lay a foundation for further study on ORF7 gene and capsid pro- tein expressed by ORF7 gene.更多还原