猪瘟病毒E2蛋白A/D抗原域在毕赤酵母中的表达  

Secretion expression conditions optimization of E2 protein A/Dantigenic domain of hog cholera virus in Pichia pastoris

作　　者：徐学清[1] 曹瑞兵[1] 蔡梅红[1] 任雪枫[1] 周斌[1] 陈溥言[1] 

机构地区：[1]南京农业大学农业部动物疫病诊断与免疫重点开放实验室,江苏南京210095

出　　处：《西北农林科技大学学报（自然科学版）》2005年第3期11-15,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家863高技术发展计划项目(2001AA249012)

摘　　要：　对含猪瘟病毒E2蛋白A/D抗原区基因插入的重组酵母菌株进行诱导表达,通过对诱导时间、重组酵母菌株、菌体密度、培养液pH、甲醇剂量等培养条件与表达产量关系的分析,对重组蛋白的表达条件进行了优化。优化后的条件为:28～30℃,225r/min振荡培养至BMGY中菌体OD600值为3.0～3.5时,将菌体转入相当于4倍体积BMGY,pH为6.0的BMMY培养基中培养72h,每24h加入甲醇至终浓度为总体积的0.5%～1.0%。在此优化条件下,重组蛋白的表达量可达275.38μg/mL,比较温度对重组蛋白降解率的影响后发现,培养物上清液应保存于-20℃。We induced the acquired recombinant Pichia pastoris strains with the gene encoding A/D antigenic domain of hog cholera virus E2 protein and optimized expression condition by studying the relations beween expression yield and growth conditions with different inductions time,recombinant strain,strain density,pH value and methanol dose of medium,respectively.The optimal conditions of the recombinant protein expression were:28-30 ℃,225 r/min,in BMMY medium of pH 6.0 which was 4 times volume of BMGY with OD_(600) being 3.0-3.5,was incubated 72 h and methanol was added to keep its concentration up to 0.5% between 24 h.After induction under optimal conditions,the yield of recombinant protein could be added up to 275.38 μg/mL.After comparing the effects of different convervation temperature on lysis of recombinant protein,it was found that the supernatant should be conserved in -20 ℃ and these jobs had provided a good basis for large-scaled ferment and application of recombinant protein.

关 键 词：巴斯德毕赤酵母 猪瘟病毒E2蛋白 A/D抗原区 表达条件 条件优化 

分 类 号：S852.651[农业科学—基础兽医学]