大肠杆菌hCG核酸疫苗质粒扩增营养条件研究  

Study on Nutritional Conditions of Plasmid Amplification of hCG DNA Vaccine in Escherichia coli

作　　者：赵雪梅[1] 夏杰[1] 陆兵[1] 徐殿胜[1] 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

出　　处：《化学与生物工程》2010年第1期63-66,共4页Chemistry & Bioengineering

摘　　要：采用摇瓶实验考察了营养条件对大肠杆菌hCG核酸疫苗质粒含量的影响。结果表明,甘油是质粒DNA生产的最佳碳源;C/N比对质粒DNA含量影响明显。向改良的PDM培养基中添加3 g.L-1的柠檬酸钠,可以降低糖酵解途径的代谢通量,使质粒含量达到11.73 mg.L-1,提高了20%。氟化钠是糖酵解途径的抑制剂,在菌体生长8 h后,向改良的PDM培养基中添加浓度为120 mg.L-1的氟化钠,培养后质粒DNA含量为12.09 mg.L-1,提高了29%。进一步研究发现乙醇对质粒DNA含量影响显著,在改良的PDM培养基中添加2 g.L-1的乙醇,可使质粒DNA含量达到12.94 mg.L-1,提高了32%。In this paper the effects of nutritional conditions on the production of plasmid of hCG DNA vaccine in E. coli on shaking-flask scale was investigated. The results indicated that glycerol was the most suitable carbon source for plasmid production, and the plasmid DNA yield was Significantly influenced by the C/N ratio. Adding 3 g ·L^-1 sodium citrate to the modified PDM m＇edium could reduce the metabolic fluxes of glycolysis pathway, which made the plasmid DNA yield reach up to 11.73 mg·L^-1, increased by 20%. Sodium fluoride was the inhibitor of glycolysis pathway,therefore 120 mg ·L^-1 sodium fluoride was added to the modified PDM medium after 8 h of incubation,and the final plamid DNA yield was 12.09 mg·L^-1 ,increased by 29%. Further study showed that ethanol had significant influence on plasmid DNA yield. Adding 2 g ·L^-1 ethanol to the modified PDM medium, the plasmid DNA yield could reach up to 12.94 mg·L^-1 , increased by 32%.

关 键 词：营养 质粒DNA 柠檬酸钠 氟化钠 乙醇 

分 类 号：Q789[生物学—分子生物学]