REK重组大肠杆菌的发酵优化
毕业论文关键词：发酵，基因工程菌，优化
Fermentation Optimization for the Recombinant Escherichia coli with Enterokinase gene
【Abstract】：Genetically engineered bacteria are genetically engineered to construct the recombinant plasmid, and thus to achieve the stable heredity and the dominant strains.This strain has the advantages of targeting, easy to culture, and easy to breed, and it can express certain products, etc... How to cultivate genetically engineering bacteria, accumulate the target product is the ultimate goal of the experiment, but also the difficulty and the key of the experiment control process.. Through the experimental verification of gene engineering bacteria, as well as in the single condition (such as temperature, feeding rate), comparison of the growth of Escherichia coli. The results showed that Escherichia coli were the most optimum fermentation conditions. The process is as follows: (1) recombinant bacteria were extracted by plasmid, and analyzed by enzyme digestion and electropHoresis, and the recombinant E. coli was selected to construct the recombinant E. coli. (2) By changing the conditions of the cultivation, optimizing the optimum conditions for its optimum growth. (3) The right induction, the maximum of the purpose of the gene to promote the existence and expression of gene. (3) The centrifugal collection is prepared for further purification. This experiment through optimization of the growth conditions can be the greatest degree of play of gene engineering bacteria advantage, but also to maximize the efficiency of fermentation production, among them a few single factors were optimized, the preliminary conclusions can be drawn: under temperature of 37℃, inoculated with 2 ml as secondary seed volume of activation, filling material in pH 6.9 and induced pH 7.0, high dissolved oxygen induced 6h, 2h induced by low dissolved oxygen, engineering bacteria can not only achieve maximum relative yield of cell and on the expression of exogenous gene is also more favorable.
Keywords: Fermentation, Gene engineering bacteria, Optimization
目录
引言 1
1. REK大肠杆菌的鉴定 2
1.1 仪器和试剂 2
1.2 REK重组质粒提取流程 3
1.3 REK重组质粒酶切流程（70ul体系）： 4
1.4 REK 重组大肠杆菌质粒电泳: 4
2.REK大肠杆菌的发酵培养 4
2.1仪器和试剂 4
2.2发酵前准备： 7
2.3发酵操作流程 7
2.4 收集菌种 8
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