猪流行性腹泻病毒S1基因在毕赤酵母中的表达  

Expression of Porcine Epizootic Diarrhea Virus S1 Gene in Pichia pastoris

作　　者：谭博敏 余希尧[1] 钟泽民[1] 蒋志琼[1] 黄毓茂[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642

出　　处：《中国畜牧兽医》2014年第6期64-67,共4页China Animal Husbandry & Veterinary Medicine

摘　　要：为了研制一种能够高效表达猪流行性腹泻病毒S1主要抗原表位基因的新型疫苗,将S1主要抗原表位基因整合到酵母表达载体pPICZαC中,构建了重组质粒pPICZαC-S1,对重组质粒经SacⅠ酶切线性化后通过电转化法转化到毕赤酵母X-33中,经ZeocinTM抗性筛选后得到阳性转化子(pPICZαC-S1),转化子接种培养基后用甲醇进行诱导,并对诱导结果进行SDS-PAGE分析和Western blotting鉴定。结果表明,纤突蛋白主要抗原表位在毕赤酵母中成功分泌表达,表达的蛋白分子质量为42ku,且能够与PEDV阳性血清特异性结合。To develop a new vaccines can efficiently express the porcine epizootic diarrhea virus S1 antigen epitope genes mainly,the recombinant plasmid pPICZaC-S1 was constructed by integrat S1 antigen epitope genes mainly into yeast expression vector pPICZaC and transformed into Pichia pastoris X33 by electroporation after linearized by Sac Ⅰ restriction enzymes. The positive transformants （pPICZaC-S1 X33） were screened by Zeocin^TM. The transformants were inoculated into culture medium and induced with methanol. The result was demonstrated by SDS-PAGE and Western blotting, and showed the mainly antigen epitope of spike protein was secreted successfully expression in Pichia pastoris. Expression of the protein molecular weight for 42 ku, and the ability to combine with PEDV positive serum specificity.

关 键 词：猪流行性腹泻病毒 抗原表位 分泌表达 毕赤酵母 

分 类 号：Q786[生物学—分子生物学]