论文半纤维素降解菌的序列测定解说.docVIP

半纤维素降解菌DJ02 16s rDNA的序列测定 和系统进化分析 生： 指导教师： 摘 要： 为研究半纤维素降解菌DJ02在分类学上的种属关系，通过对菌株进行形态观察，革兰氏染色，生理生化反应，提取总基因组DNA。利用通用引物对16S rDNA序列进行PCR扩增，并对扩增到的目标片段进行测序，将测序结果与NCBI上已知菌种的16S rDNA 序列进行BLAST 对比，构建系统进化树进行分析。结果表明，该菌为革兰氏阴性杆菌，V-P反应呈阴性，甲基红和吲哚试验呈阳性，能分解葡萄糖、蔗糖和少量乳糖，产酸不产气。扩增片段长度为1502bp，与预期的1500bp相符。核苷酸相似性分析发现，菌株DJ02与NCBI基因库中的不动杆菌属的菌株聚为一簇，并且与Uncultured Acinetobacter sp. clone GI5-004-C08菌株在同一分支中，与不动杆菌属Uncultured Acinetobacter sp. clone GI5-004-C08菌株16S rDNA核苷酸序列相似性最高为99%。结论表明菌株DJ02属于不动杆菌属，命名为Acinetobacter sp. DJ02。不动杆菌属；16S rDNA；系统发育分析；鉴定 Sequencing and Phylogenetic Analysis of the 16S rDNA of Hemicelluloytic Bacterial Community DJ02 Undergraduate: Deng li Supervisor: Jiang Li-chun Abstract: In order to identify the species relationship of Hemicelluloytic Bacterial Community DJ02. Firstly, the morphology of the Bacteria was observed, then the Gram Stain, physiological and biochemical responses and the extraction of total genomic DNA had been done. After that, the target 16S rDNA sequence was amplified by using the general primers, and this sequence was analysised by the NCBI with BLAST data base. As a result, the Bacteria was Gram-negative bacilli, the V-P reaction was negative, Methyl red and indole test were positive; And the Bacteria could break down the glucose, sucrose and a small amount of lactose, producing acid no gas. 1500bp desired fragment was amplified and nucleotide sequence of 1502bp was obtained from the bacteria. The sequence had the highest similarity of 99％ with 16S rDNA of GI5-004-C08 strain of Uncultured Acinetobacter sp. clone. The result showed that the strain DJ02 and Acinetobacter strain formed a separate and distinct phylogenetic branch which revealed that the strain DJ02 is Acinetobacter species, it was named Acinetobacter sp. DJ02. Key words:Acinetobacter; 16S rDNA; phylogenetic analysis; identification