摘要
本研究对影响FGF21(L^(59)R)突变体基因工程菌发酵条件的因素进行了优化。采用正交试验确定FGF21(L^(59)R)突变体基因工程菌的最佳LB培养基配方,利用SDS-PAGE电泳检测不同发酵条件对FGF21(L^(59)R)突变体蛋白表达情况。LB培养基最佳配比(g/L):蛋白胨11,酵母粉6,氯化钠10,葡萄糖1;在此基础上优化基因工程菌发酵条件,确定LB培养基p H 6.5~7.2,装液量(溶解氧)20%,菌体密度(A_(600))1.0,IPTG浓度0.6 mmol/L,37℃条件下诱导5 h,突变体蛋白的表达量由优化前的12%提高至35%。结果表明,培养基配方、p H、装液量(溶解氧)、菌体密度(A_(600))、IPTG浓度、温度、诱导时间均对FGF21(L^(59)R)突变体基因工程菌表达量有影响。
In this study, we optimized the factors of influencing the fermentation conditions of FGF2 1 （L59R） mu- tant gene engineering bacteria. We determined the best LB culture medium formula of FGF2 1 （L59R） mutant genetic engineering bacteria by orthogonal test. We detected the expression of protein of FGF2 1 （L59R） mutant by SDS- PAGE electrophoresis. The optimal ratios of LB medium components （g/L） were peptone 11, yeast powder 6, sodium chloride 10, and glucose 1. We optimized the fermentation conditions on the above optimal medium and found that the expression value of mutant protein increased from 12% to 35% when the pH of LB medium was 6.5-7.2, liquid volume in flask was 20%, growth density of the strain （A600） was 1.0, IPTG concentration was 0.6 retool/L, and inducing time was 5 h at 37℃. The results showed that medium components, pH, liquid volume, growth density of the strain （A600）, IPTG concentration, inducing time and inducing temperature would affect the expression value of FGF2 1 （L59R） mutant genetic engineering bacteria.
作者
陈秋池
王会岩
朱文赫
董媛
王宁宁
李艳
Chen Qiuchi Wang Huiyan Zhu Wenhe Dong Yuan Wang Ningning Li Yan(School of Laboratory, Jilin Medical College, Jilin, 132013)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2016年第9期2398-2403,共6页
Genomics and Applied Biology
基金
国家自然科学基金项目(81273421)
吉林省教育厅项目2014第361号共同资助
