PCR扩增技术连云港盐田嗜盐细菌多样性研究
毕业论文关键词：盐田；嗜盐细菌；454高通量测序；16S rRNA基因；多样性
The Diversity of Halophilic bacterial in the Lianyungang salt field
Abstract：To obtain the community structure and persity of the halophilic bacteria in Lianyungang sail field,we use 454 high-throughput 16S rRNA sequencing.6272 valid sequence and 5558 optimized sequence and 420 OTUs were obtained finally. Compared with archaea, bacteria showed higher microbial richness and persity.The bacterial community was dominated by the classes Cytophagia in Bacteroidetes phylum and Gammaproteobacteria in Proteobacteria phylum.The study of persities of halobacteria in Lianyungang sail pond can not only help us to know the general features of local bittern at the level of microorganism,to improve and develop biotechnology in salt industrial manufacture.It can also enrich and protect the species resource of extremophiles in our country,and provide important reference for the exploitation of new species.
Keywords: salt field; Halophilic bacterial; 454 high-throughput sequencing; 16S rRNA; Diversity.
目录
引言 1
1材料与方法 2
1.1供试材料 2
1.1.1水样采集 2
1.1.2 采样地点介绍 2
1.2 实验流程与方法 2
1.2.1 宏基因组DNA提取 2
1.2.2 设计并合成引物接头 2
1.2.3 PCR扩增及产物纯化 3
1.2.4 PCR产物定量及均一化——荧光定量法 3
1.2.5 454高通量测序 3
1.3原始数据处理 3
1.3.1有效序列数据处理 3
1.3.2优化序列数据处理 3
1.4生物信息学分析 4
1.4.1 OTU生成 4
1.4.2分类学分析 4
1.5相关指数与曲线说明 4
1.5.1 稀释性曲线 4
1.5.2 多样性指数 4
2结果与分析 4
2.1 454高通量测序结果 4
2.2丰度、多样性数据分析 5
2.2.1 OTU划分 5
2.2.2菌群丰度 5
2.2.3菌群多样性 5
2.2.4测序深度 5
2.3 菌群结构分析 6
2.3.1 门（phylum）水平上菌群结构 6
2.3.2 纲（class）水平上菌群结构 7