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摘要：采用现代分子生物技术和基因重组技术，敲除野生菌恶臭假单胞菌菌KT2440中ATP合成酶中α亚基的合成基因，构建出工程菌KT2440 datpA。同时，将ATP水解酶重组到质粒pBBR1MCS5上构建重组质粒pBBR1MCS5-KT-atpAGD，然后转化入野生菌KT2440。将野生菌与工程菌同时培养在以芳烃化合物为唯一碳源的MM培养基中，比较野生菌与工程菌的生命活动及其降解芳烃化合物的速度，探讨敲除基因atpA和构建的重组质粒对KT2440在降解芳烃化合物的过程中有何影响。结果表明atpA基因的敲除没有明显影响，而重组质粒转化进KT2440对其生命活动及其降解芳烃化合物均有明显的影响。63947
毕业论文关键词 KT2440; 基因敲除; 重组质粒; 芳烃化合物
毕业设计论文外文摘要
Title Comparison the ability of degradation of pollutants Of pseudomonas putida and mutant strain
Abstract Using modern molecular biotechnology and gene recombination technique, knockout the gene synthesizing α subunit of ATP synthase in wild strain Pseudomonas putida KT2440 to construct gene engineering strain KT2440 datpA. And at the same time, reorganize ATP hydrolase to plasmid pBBR1MCS5 to form pBBR1MCS5-KT-atpAGD transformed to the wild strain KT2440. Cultivate KT2440 and engineering bacteria in MM medium with aromatic compounds as the sole carbon source.Then, compare the life activity and the degradation speed of aromatic compounds between wild strain and engineering strain, and explore the effect of the knockout of atpA gene and construction of recombinant plasmid pBBR1MCS5-KT-atpAGD in the process of KT2440 degrading aromatic compounds. The results showed that the knockout of the atpA gene had no obvious effect, and transforming the recombinant plasmid pBBR1MCS5-KT-atpAGD into the KT2440 has the obvious effect on the life activity and the degradation ability of aromatic compounds.
Keywords KT2440; gene knock-out; recombinant plasmid; aromatic compound
1 引言 1
1.1 微生物对有机农药的降解 1
1.1.1 降解有机磷农药的主要微生物菌群 1
1.1.2 微生物降解有机农药在国内的研究状况 2
1.2 微生物对芳烃化合物的降解 3
1.2.1 微生物降解芳烃化合物的机理 3
1.2.1.1 国内的微生物代谢芳烃化合物的研究状况 4
1.2.1.2 微生物降解氯代硝基苯类化合物的途径和机理研究 5
1.2.1.3 氯代硝基苯类化合物在生物修复上的研究应用 5
1.3 芳烃化合物的降解基因研究 5
1.3.1 降解基因的克隆研究 6
1.3.2 芳香烃降解基因工程菌的构建研究 6
1.3.3 基因工程载体的构建 7
1.3.4 ATP合成酶的结构和功能