摘要
利用果蝇S2细胞表达鸡传染性囊病病毒(IBDV)超强毒株VP2蛋白,并检测其抗原活性。通过扩增IBDV VP2蛋白的编码基因,与果蝇S2细胞表达载体pMT/BiP/V5/HisA连接构建重组表达载体pMT/BiP/V5/HisA-VP2,将重组表达载体与筛选质粒pCoBlast共转染果蝇S2细胞,表达VP2融合蛋白后纯化目的蛋白,并对表达产物进行抗原性分析。Western-blotting分析表明,融合蛋白相对分子质量为49 kDa,该融合蛋白具有与VP2单抗良好的结合能力。结论 IBDV VP2融合蛋白能在果蝇S2细胞中进行有效地表达,并且能分泌到上清中,融合蛋白具有良好的抗原性,可作为诊断抗原用于该病的诊断检测。
To express very virulent infectious bursal disease virus （vvIBDV） strain LX VP2 protein by Drosophila s2 cell expres- sion system and to further analyze the antigenicity of the recombinant protein, the VP2 gene from vvIBDV strain LX was amplified by reverse transcription （RT）-PCR and cloned into Drosophila $2 cell expression vector pMT/BiP/V5/HisA to obtain a recombinant ex- pression vector pMT/BiP/V5/HisA-VP2. The plasmid from the pMT/BiP/V5/HisA-VP2 vector aiad screening plasmid pCoBlast were cotransfected into $2 cells to generate VP2 fusion protein. Then the expressed protein was purified and its antigenicity was analyzed. Results show that a specific and simple band with a size of about 49 kDa was observed as examined by Western Blot using a monoclo- nal antibody raised against VP2 protein indicating that IBDV VP2 protein can be effectively expressed in $2 cells followed by secret- ing into the supernatant with a good antigenicity, and it can used as an antigen for diagnosis.
出处
《中国兽医杂志》
CAS
北大核心
2014年第2期10-13,共4页
Chinese Journal of Veterinary Medicine
基金
国家现代农业产业技术体系(CARS-42)
