【作者】 蔡正求；

【导师】 宋未；

【作者基本信息】 首都师范大学 ， 植物学， 2004， 硕士

【摘要】 核酸二级数据库是生物信息学研究的重要领域，对生命科学的研究和发展起重要作用。目前，国际核酸序列公共数据库中存在大量的细菌16S rDNA序列，本研究从国际核酸数据库中收集了所有已知细菌的16S rDNA序列，根据这些序列设计了大约200种细菌的种特异性的16S rDNA寡核苷酸探针，将结果存入数据库，开发基于WEB的查询系统，查询结果形象直观，为设计细菌的种特异性寡核苷酸探针提供参考和帮助。 在植物根际存在大量的促生细菌，对这些细菌进行快速的分类地位检测对研究植物与微生物的相互作用和微生物接种剂应用的有效性有重要意义。本研究选择植物根际常见促生细菌，即巴西固氮螺菌(Azospirillum brasilense)、生脂固氮螺菌(Azospirillum lipoferum)、蜡状芽孢杆菌(Bacillus cereus)、巨大芽孢杆菌(Bacillus megaterium)、短小芽孢杆菌(Bacillus pumilus)、球形芽孢杆菌(Bacillus sphaericus)、枯草芽孢杆菌(Bacillus subtilis)、凝结芽孢杆菌(Bacillus coagulans)、多粘类芽孢杆菌(paenibacillus polymyxa)、唐菖蒲假单孢菌(Burkholderia gladioli)，土壤杆菌属(Agrobacterium)、产碱菌属(Alcaligenes)、节杆菌属(Arthrobacter)、慢生根瘤菌属(Bradyrhizobium)、伯克霍尔德氏菌(Burkholderia)、黄杆菌属(Flavobacterium)、假单孢菌属(Pseudomonas)、根瘤菌属(Rhizobium)、链霉菌属(Streptomyces)和科肠杆菌科(Enterobacteriaceae)，分别设计这些不同分类水平(种、属、科)细菌的探针，将其固定在特殊处理的载玻片上，用相关的细菌对这些探针进行筛选，选择特异性较好的探针，将用于对未知样品进行检测。结果表明大部分探针都有较好的杂交信号。更多还原

【Abstract】 Secondary database that play an important role in the research and development of biology is a vital research subject in the field of bioinformatics. At present, there are enormous 16S rDNA sequences of bacteria available in the Genbank. In this research all 16S rDNA sequences of bacteria are collected and the about two hundred species-specific oligonucleotide probes for various bacteria according to 16S rDNA have been designed and stored into database. And a user-friendly search system based on computer network has been constructed. This secondary database could help users to design oligonucleotide probes.There are a number of plant growth-promoting rhizobacteria in the rhizosphere of plants and it is meaningful for detecting, classifying and identifying these bacteria to research the interaction of plants and microorganisms and the efficiency of the microbial inoculums. In this research, familiar plant growth梡romoting rhizobacteria are selected to design respectively oligonucleotide probes which target for different classification levels (species, genera and family) of bacteria and will be fixed to specific slides and then relative bacteria are used to select specific probes which would be applied to detect unknown examples. These rhizobacteria include Azospirillum brasilense, Azospirillum lipoferum, Bacillus cereus , Bacillus megaterium , Bacillus pumilus , Bacillus sphaericus , Bacillus subtil is , Bacillus coagulans , paenibacillus polymyxa , Burkholderia gladioli , Agrobacterium , Alcaligenes , Arthrobacter, Bradyrhizobium, Burkholderia, Flavobacterium, Pseudomonas, Rhizobium, Streptomyces and Enterobacteriaceae. The results show that it is successful for most probes to produce positive signals of hybridization.更多还原