中度噬盐细菌的分子鉴定531.doc

7株中度嗜盐细菌的分子鉴定 刘亮 指导老师：温洪宇 （江苏师范大学生命科学学院，江苏 徐州 221116） 摘要： 【目的】通过16S rRNA基因的扩增，对七株中度嗜盐菌进行分子鉴定和分类学地位的研究。【方法】对7株4℃冷冻保藏的中度嗜盐细菌进行活化，对其提取DNA后测序，按系统进化树的聚类关系，结合基于16S rRNA基因系统发育分析的方法[3]来研究七株中度嗜盐细菌的分类学地位。【结果】结果表明QD1与Halomonas venusta DSM 4743（T）相似度为98.456%，QD6与Virgibacillus litoralis JSM 089168（T）相似度为95.909%，QD11与Marinobacter flavimaris SW-145（T）相似度为100.00%，QD12与Halobacillus trueperi DSM 10404（T）相似度为99.868%，WY17与Virgibacillus litoralis JSM 089168（T）相似度为95.976%，3T与Pontibacillus chungwhensis BH030062（T）相似度为99.719%，9与Cobetia marina DSM 4741（T）相似度为99.800%。【结论】7株中度嗜盐菌属于菌群Halomonas venusta QD1，Virgibacillus litoralis QD6，Marinobacter flavimaris QD11，Halobacillus trueperi QD12，Virgibacillus litoralis WY17，Pontibacillus chungwhensis 3T，Cobetia marina 9。 关键词：中度嗜盐菌；分子鉴定；16SrRNA；物种分类；进化树 Molecular identification of seven moderately halophilic bacteria Liu Liang supervision:Wen hongyu (School of Life Sciences,Jiangsu Normal University, Xuzhou, Jiangsu ,221116,China) Abstract: [Objective] The molecular identification and specices classification of seven moderately halophilic bacterias which identified by 16S rRNA gene sequence analysis.[Methods] Reactivate seven moderately halophilic bacterias,3T，9，QD1，QD6，QD11，QD12 and WY17 were numbered.Each bacterial genomic DNA was extracted by genomic DNA extraction kit and 16S rRNA gene were sequenced subsequently.Analysis the species classification by phylogenetic tree construction based on 16S rRNA gene[3].[Results] The result showed that seven moderately halophilic bacterias were Halomonas venusta QD1，Virgibacillus litoralis QD6，Marinobacter flavimaris QD11，Halobacillus trueperi QD12，Virgibacillus litoralis WY17，Pontibacillus chungwhensis 3T，Cobetia marina 9。 Key word: Moderately halophilic bacteria; Molecular identification; 16SrRNA; Species classification；Phylogenetic tree 目录 目录 3 引言 1 1材料与方法 2 1.1试验材料 2 1.1.1 待活化菌株 2 1.1.2 培养基 2 1.1.3 试剂与仪器设备 2 1.2试验方法 2 1.2.1 菌株活化 2 1.2.2 对七株中度嗜盐菌分别进行革兰氏染色 3 1.2.3 对单株中度嗜盐菌进行D