核酸疫苗载体pVAX-44质粒高拷贝研究  

Study on Plasmid Amplification of DNA Vaccine Vector pVAX-44

作　　者：丁霞[1] 陆兵[1] 夏杰[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》2008年第6期56-59,62,共5页Chemistry & Bioengineering

基　　金：上海市科委生物医药重点科技攻关项目(064319035)

摘　　要：研究了质粒复制的前体物质、蛋白质合成抑制剂及金属离子对核酸疫苗载体pVAX-44质粒拷贝数的影响。结果表明,在改良的LB培养基中分别添加谷氨酰胺、甘氨酸、天门冬氨酸和Fe3+时,单位菌体质粒含量分别达到5.88mg.g-1、7.15 mg.g-1、5.93 mg.g-1、7.38 mg.g-1,同时添加这几种物质时,质粒拷贝数可以得到进一步扩增。均匀设计实验结果表明,改良的LB培养基中含有1.6 g.L-1甘氨酸、1.0 g.L-1天门冬氨酸、0.2 g.L-1谷氨酰胺、0.4mmol.L-1Fe3+时,单位菌体质粒含量可达到9.11 mg.g-1,明显高于单独添加这几种物质时的质粒拷贝数。在此基础上加入0.1 g.L-1链霉素抑制蛋白质合成,单位菌体质粒含量可达到9.53 mg.g-1,较对照提高了56.25%。The effects of plasmid precursor,protein synthesizing inhibitors and metal ions on the amplification of pVAX-44 （DNA vaccine vector） were studied. The results showed that plasmid per DCW amounted up to 5.88 mg · g^-1,7.15 mg · g^-1,5.93 mg· g^-1,7.38 mg·g^-1 individually through adding Gln,Gly,Asp and Fea＋ into improved LB medium,respectively. Plasmid amplification increased dramatically through adding them into improved LB medium simultaneously. Uniform design experiments results showed that plasmid per DCW amounted up to 9.11 mg · g^-1 as the improved LB medium containing 1.6 g · L^-1 Gly,1.0 g · L^-1 Asp,0.2 g · L^-1 Gln, and 0.4 mmol · L^-1 Fe^3＋ ,higher than that of adding them individually. Based on these,0.1 g · L^-1 streptomycin as protein synthesizing inhibitor was added and the plasmid amplification reached 9.53 mg · g^-1, increasing by 56.25% after optimization.

关 键 词：质粒 拷贝数 核酸疫苗 均匀设计实验 

分 类 号：Q789[生物学—分子生物学]