摘要
目的:筛选体外高表达商陆抗病毒蛋白的毕赤酵母重组菌株,并摸索其适合的发酵条件。方法:通过电击转化将含有商陆抗病毒蛋白(pokeweed antiviral proteins,PAP)基因的分泌型表达载体PIC9K-P导入到毕赤酵母(Pachia pastoris)GS115菌株中。利用免疫印迹法筛选表达量较高的转化子。在摇瓶发酵水平对重组菌株产PAP条件进行初步研究。结果:高表达PAP的重组菌株在发酵时间为96h,10g/L的甲醇浓度,培养基pH值为6.0～6.4时PAP的表达量较高。结论:免疫印迹法适合用于毕赤酵母高表达重组菌株的筛选,重组毕赤酵母的PAP表达量可高达30mg/L。
Objective：In this experiment,high expression transformants of PAP in Pichia pastoris and its suitable fermentation conditions were studied.Method：A secreted expression vector pPIC9K-P including the pokeweed antiviral protein （ PAP） gene was transformed electrically into Pichia pastoris GS115.High expression transformants was screened by immunoblotting test.The optimal fermentation conditions for producing PAP in shake-flask cultivation was investigated.Result：96 hours culture time,10g/L methanol and pH 6.0-6.4 of culture condition will induced high expression of PAP.Conclusion：Immunoblotting test is suitable for screening high expression transformants and the expression of PAP in reconbinant Pichia pastoris is over 30mg/L.
出处
《生物技术》
CAS
CSCD
北大核心
2010年第4期62-64,共3页
Biotechnology
关键词
商陆抗病毒蛋白
毕赤酵母
高表达重组菌株
发酵条件
pokeweed antiviral protein
PAP
Pichia pastoris
high expression transformants
fermentation conditions
