近海细菌hzo基因及古菌amoA基因多样研究毕业论文.doc

中国石油大学（华东）毕业设计（论文） 近海基因及古菌基因多样性研究 摘 要 胶州湾和海南岛海域优美的自然环境与独特的地质演化进程吸引了众多中外科学家的关注。这种复杂的地质演化过程必然也伴随着生物的不断进化。 本实验中涉及的hzo和amoA基因序列可用于物种多样性的分析，相对于比较DNA所有序列，更简便并有代表性。本实验通过对胶州湾A5站位微生物hzo基因以及海南岛海域E505站位amoA基因的PCR扩增，转入感受态大肠杆菌，酶切分析，测序比功能比对等一系列步骤，建立站位的微生物系统树。 本实验所采集的样品来自于胶州湾和海南岛海域，实验采用对沉积物进行hzo和amoA基因文库构建进一步测序分析进化关系的方法。通过建立的系统发育树可以对两个站位的微生物多样性进行评估。本次实验证明胶州湾A5站位hzo基因多样性较少而海南岛海域E505站位amoA基因有较高多样性。 关键词：厌氧氨氧化；hzo基因；amoA基因 abstract The beautiful natural sceneries and the unique process of geological evolution of Jiaozhou Bay and Hainan Island waters are attracting more and more Chinese and foreign scientists. The complex geological evolution may go with continuous evolution of living beings. This study analyzed the archaeal and bacterial diversity by the construction of gene sequences that can be applied to the analysis of species diversity. In this experiment, sediment DNA from Jiaozhou Bay and Hainan Island coastal areas was extracted; hzo genes and amoA genes were amplified through PCR with degenerate primers justified by former researches, inserted into plasmids, which were then transformed with competent E.coli cells, analyzed by RFLP, and sequenced. After series of procedures, phylogenetic trees based on hzo and amoA genes were constructed, which is a good indication of microbe diversity. The sediment samples used in this study were collected from the Jiaozhou Bay and Hainan Island Waters. This study analyzed the archaeal and bacterial diversity by the construction of hzo and amoA gene libraries. From the two phylogenetic trees conducted, we can see hzo genes of staion A5 of Jiaozhou Bay has low diversity while amoA genes of station E505 of Hainan Island has high diversity. Keywords: Anammox; hzo genes; amoA genes目录 第1章 前言 1 1.1 胶州湾生态环境 1 1.2 厌氧氨氧化细菌 1 1.3 该实验涉及的的分子生物学技术 3 1.3.1 PCR技术 3 1.3.2 限制性酶切 6 1.3.3 琼脂糖凝胶电泳 7 1.4 本文的研究目的及意义 8 1.4.1 本文的研究目的 8 1.4.2 本文的研究意义 9 第2章 实验部分 10 2.1 材料及仪器 10 2.1.1 胶州湾的地理位置及站位来源 10 2.1.2 实验药品 10 2.1.3培养基 11 2.1.4主要仪器设备 11 2.2 实验步骤