摘要
以PRRSV细胞培养物为模板,经RT-PCR扩增得到ORF5/ORF6基因,该产物和pIRES真核表达载体分别经双酶切,连接构建pIRES-ORF5/ORF6。经PCR、双酶切鉴定和测序证明成功构建了真核表达载体pIRES-ORF5/ORF6。测得的ORF5/ORF6序列与VR-2332株的氨基酸同源性分别为99%和100%,属北美洲型。
ORF5 and ORF6 genes of PRRSV strain SD2 were amplified by RT-PCR, pIRES-ORF6 was constructed by the ligation of pIRES and ORF6 digested with Nhe Ⅰ and EcoR Ⅰ pIRES-ORF5ORF6 was constructed by the ligation of pIRES-ORF6 and ORF5 digested with Xba Ⅰ and Not Ⅰ We succeeded in constructing pIRES-ORF5ORF6 identified by PCR, double digestion and nucleotide sequencing. The homologus of deduced amino acids sequences was 99% and 100% of ORF5 ORF6 compared to VR-2332 and belonged to America type. A recombinant eukaryotic vector, which expressed two proteins, was constructed and could provided a kind of DNA vaccine. This study provided some basic data for further development of PRRS subunit vaccine .
出处
《动物医学进展》
CSCD
2006年第10期78-81,共4页
Progress In Veterinary Medicine
基金
山东省自然基金重大项目(Z2000D03)
