嗜热菌分离筛选及分子分类初探  

Isolation and Identification of Thermophilic Microorganisms

作　　者：阮灵伟[1] 刘欣[2] 杨海杰[3] 李钫[2] 

机构地区：[1]厦门大学生命科学学院,福建厦门361005 [2]国家海洋局第三海洋研究所海洋生物遗传资源重点实验室,福建厦门361005 [3]厦门大学海洋与环境学院,福建厦门361005

出　　处：《厦门大学学报（自然科学版）》2006年第2期276-279,共4页Journal of Xiamen University：Natural Science

基　　金：中国大洋矿产资源研究开发协会基金(DY105-02-04-05);863-中国高技术研究发展计划基金(2004AA621010)资助

摘　　要：从厦门周边地区温泉采集得到样品,在75℃和80℃的温度下进行培养并分离得到60株嗜热菌.为了从大量的菌株中筛选出不同的菌株,有效地简化筛选过程,应用了细菌全蛋白SDS-PAGE电泳、RAPD和16S-23S rDNA间隔区比较分析等分子生物学方法从中筛选得到7株种属不同的嗜热菌,并进一步运用16S rDNA序列分析鉴定其种属关系,为深入研究奠定了基础.Thermophilic bacteria is an important biological resource and plays an important part in many fields of biotechnology. Traditionally, taxon of bacteria is usually based on the phenotype that is time-consuming. So we applied some molecular methods to facilitate the process of classification. The samples were collected from a hot spring around Xiamen near the sea and the thermophilic bacteria were cultured at 75℃ and 80℃. Then the single clony of bacteria was isolated and 60 strains were obtained in total. In order to facilitate the process of classification from a large number strains of bacteria, bacterial populations were identified by the three methods of molecular biology,including SDS-PAGE electrophoresis of whole bacterial cell protein, RAPD and amplification of 16S-23S rDNA spacer regions. By combining these three methods,we had isolated 7 strians of bacteria that belong to different genus among 60 strains. They showed different electrophoresis fingerprints and can be easily distinguished from each other among 60 strains of bacteria. These 7 different strains were further confirmed by sequencing the 16S rDNA after amplification by PCR. Then the sequences were uploaded to NCBI to compare with other bacteria that had been found before. On the basis of these datas,phylogenetic tree of the 7 strains of bacteria was constructed and their relationship can be determined. These results provided a reliable basis for the further study on these thermophilic bacteria. Furthermore,combination of these 3 molecular methods could be used as a simple way to classify bacterial strains.

关 键 词：嗜热菌 全蛋白 RAPD 16S-23S rDNA间隔区 

分 类 号：Q933[生物学—微生物学]