摘要
【目的】获得鸡α干扰素重组酵母菌诱导表达的最佳诱导时间和甲醇的体积分数,并测定诱导表达产物对鸡马立克氏病毒(MDV)和鸡新城疫病毒(NDV)的抑制能力。【方法】挑取2个鸡α干扰素重组酵母菌单菌落至BMGY培养基中培养,待菌液OD600为4左右时,分别转接至BMMY培养液诱导,1瓶每隔12 h补加1次体积分数0.5%甲醇,另1瓶每隔12 h补加1次体积分数1%甲醇,于24,48,72 h收集样品,备SDS-PAGE检测用。根据不同时间(24,48,72,96 h)诱导上清液表达量的差异,留用表达量最高时段的上清液,经初步纯化后,利用鸡胚成纤维细胞(CEF),分别测定重组酵母鸡α干扰素抑制MDV及NDV的能力。【结果】鸡α干扰素重组酵母菌在每隔12 h补加1次体积分数1%甲醇诱导48 h时,或每隔12 h补加1次体积分数0.5%甲醇诱导72 h时,重组酵母鸡α干扰素表达量均达到最高;且诱导上清液具有较好地抑制MDV、NDV的能力。【结论】获得了鸡α干扰素重组酵母菌小量发酵的最佳条件;获得的重组酵母鸡α干扰素对MDV和NDV有明显的抑制作用。
【Objective】 The research was to find the optimal conditions for induction and volume fraction of the recombinant ChIFN-α yeast and study its activity of inhibiting Markers Disease virus(MDV) and Newcastle Disease virus(NDV).【Method】 Two recombinants were induced at different time and different concentrations of methanol,the production of recombinant ChIFN-α was detected by SDS-PAGE.The antivirus activity to MDV and NDV of the ChIFN-α was detected in the chicken embryonic fibroblast(CEF).【Result】 The optimal...
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2010年第9期37-41,共5页
Journal of Northwest A&F University(Natural Science Edition)
基金
江苏大学高级人才专项资助项目(1283000152)
