摘要
为研究羊口疮病毒(ORFV)ORF129基因编码的锚蛋白调节宿主天然免疫应答的作用机制,利用PCR技术对ORFV/QH01/2010分离株ORF129基因进行扩增,并连接到pGEM-T easy载体,构建pGEM-ORF129重组质粒,双酶切后与pEGFP-N1真核表达载体连接,构建pEGFP-ORFV-ORF129真核表达质粒,经酶切、PCR鉴定及测序正确后转染BHK-21细胞,观察其在细胞中的表达情况.结果表明:ORFV ORF129基因在BHK-21细胞中获得稳定表达,并为进一步研究其编码蛋白在ORFV感染过程中的分子机制奠定了基础.
For the study of the protein which encoded by orf virus ORF129 gene modulated the host na ture immune response mechanism,PCR method was used to amplify the 0RF129 gene which derived from ORFV/QH01/2010 and cloned into pGEM-T easy vector to construct the recombinant plasmid pGEM （）RF129. The target gene was double digested and cloned into the eukaryotic expression vector pEGFP-N1 to construct eukaryotic expression vector named pEGFP-ORFV- ORF129. The recombinant plasmid was transfected BHK-21cells and observed its expression after the open reading frame was identified by enzyme digestion and PCR. The result showed that the 0RF129 gene of ORFV was successfully expressed in BHK21 cells and laid the foundation for further study of the molecular mechanism of the protein encoded by ORFV during infection process.
出处
《甘肃农业大学学报》
CAS
CSCD
北大核心
2013年第5期8-13,共6页
Journal of Gansu Agricultural University
基金
国家十二五“863”课题(2011AA10A211)
甘肃省农业生物技术研究专项(GNSW-2012-16)
家畜疫病病原生物学国家重点实验室自主研究课题
