具有氨氧化功能大肠杆菌工程菌的构建-轻工技术与工程专业毕业论文.docxVIP

活性。综上所述，本研究构建了一株具有氨氧化功能的大肠杆菌工程菌，在氨 活性。 综上所述，本研究构建了一株具有氨氧化功能的大肠杆菌工程菌，在氨 氮污水处理中有一定的应用价值。 关键词：氮素污染，氨氧化细菌，amoA基因，hao基因，脱氮活性 lI 万方数据 Construction Construction of the Engineering Escherichia with Ammonia Oxidizing Function ABSTRACT Water pollution is a significant environmental problem，nitrogen is one of the main pollution comtaminants．At present，nitrification-denitrification process is used to remove nitrogen from sewage．Ammonia oxidizing bacteria(AOB)is maj or microbe in remove nitrogen which can convert ammonia nitrogen into nitrite．However，the growth rate of ammonia oxidizing bacteria is slow and it is difficult to cultivate and isolate．it iS still a limiting factor in the process of wastewater treatment．Therefore，it is that building a strain that grows faster and better will promote the development of biological denitrification technology． In this paper,the two functional genes(amoA and hao)ofAOB were cloned from the enrichment soil solution，then two genes were made to express simultaneously in the same Escherichia coli BL2 1(DE3)，the engineering E．coli with ammonia oxidizing function was constructed．The main results were as follows： (1)With the total DNA extracted from enrichment soil solution，amoA and hao sequences were amplified by polymerase chain reaction(PCR)with the specific primers of AOB．Amplified products respectively connected with pMD 1 8-T vector,after transformation of E．coli DH5a，the positive transformants were selected by colony PCR，sequenced and analyzed by Blast，two kinds of gene sequence were obtained．The result showed that the sequences of amoA and hao had 99％indentities with Nitrosomonas sp．GH22 and Nitrosomonas sp． ENI一1 1，respectively． (2)Bioinformatics was used to analyze the structure and function of by AmoA and Hao，the analysis showed AmoA encoded 276 amino acids，its molecular weight was 3 1．94 kDa．It was a kind of unstable hydrophobic protein with potential transmembrane helix regions，and the tertiary structure prediction showed that spa