Phylogenetic Analysis and Specific DNA Probe Selection of Soil Denitrifiers
| Autor: | HSIEH, HSING-CHUAN, 謝幸娟 |
|---|---|
| Rok vydání: | 1998 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 86 Phylogenetic relationship among 26 bacteria including 21 denitrifiers, 4 enterobacteria and 1 nondenitrifier were distinguished by PCR methods that rely on different amplification priming strategies: RAPDs (random amplified polymorphic DNAs) PCR, REP ( repetitive extragenic palindromic ) PCR and nosZ conserved region PCR. The PCR products by 7 random primers, 2 REP primers and 3 nosZ primers were analyzed and compared to one another. However, the bacteria we used have large diversity in taxanomy, the results of the PCR assays were not fully consistent with their classification, and we may choose appropriate primers for analysis. However primer 96 could be used to distinguish 9 strains of Alcaligenes faecalis subsp. faecalis. And some genus-specific or species-specific DNA fragments in PCR products could be selected as genetic markers for identification of these organisms. We were interested in one of the soil bacteria-Thiosphaera pantotropha which has the capacity of aerobic denitrification and heterotrophic nitrification simutaneously. Thus, the genomic DNA of T. pantotropha was digested by restriction enzymes and cloned randomly. The isolated DNA fragements were labeled by using nonradioactive label system, and hybridization with 26 soil bacteria to select specific probes for T. pantotropha were performed. However, it is easily to detect Alcaligenes sp. by DNA fragments digested with BamHI, EcoRI and HindⅢ. Besides, the probe prepared from amplifying the unconserved region of the nosZ gene of Pseudomonas stutzeri by PCR method had high specificity to Ps. stutzeri itself. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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