| Summary: | 碩士 === 國立臺灣大學 === 植物學系研究所 === 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.
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