【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。更多还原

【Abstract】 To express very virulent infectious bursal disease virus(vvIBDV) strain LX VP2 protein by Drosophila S2 cell expression 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 S2 cell expression vector pMT/BiP/V5/HisA to obtain a recombinant expression vector pMT/BiP/V5/HisA-VP2. The plasmid from the pMT/BiP/V5/HisA-VP2 vector and screening plasmid pCoBlast were cotransfected into S2 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 Westem Blot using a monoclonal antibody raised against VP2 protein indicating that IBDV VP2 protein can be effectively expressed in S2 cells followed by secreting into the supernatant with a good antigenicity,and it can used as an antigen for diagnosis.更多还原