Study Of The Establishment Of Of Actinobacteria Fish Identification And Taxonomy

One of the main lines of descent with the domain Bacteria includes a wide range of morphologically diverse organisms, most of which, on the basis of a gram-positive staining reaction, can be considered members of the division Firmacutes. The lineage - Actinobacteria class nov. comprises organisms with a DNA base composition which generally is above 50% G+C (with a few exceptions).Some species of Actinobacteria have biotechnological or medical importance. Since the discovery of Streptomycin, a large number of antibiotics have been isolated from the cultures of a wide variety of Streptomycetes and rare Actinomycetes.For increasing the chance of discovering new useful compounds from microbial, we expanded the scope of screening targets from the members of Actinomycetes to Actinobacteria. I focus on studying bacteria-like, Actinobacteria.We determinated the sensitivity of some bacteria comprising gram-negative bacteria, gram-positive low G+C bacteria, Actinobacteria and Actinomycetes against 12 kinds of antibiotics. We added 25μg/ml Nalidixic acid and 25μg/ml Aztreonam into isolation media to inhibit the other bacteria and 20μg/ml Benlete to inhibit fungi. We set up a method that has been used in isolating Actinobacteria and Actinomycetes.From the four soil samples, 64 bacteria strains were isolated. Among them, there were 54 gram-positive bacteria, which was identified by gram staining using Live Baclight? Bacteria Gram Stain Kit.We used fluorescent in situ hybridization to identify Actinobacteria. Using the PU probe suitable for all bacteria and its control probe PN, combined with the probe PHGC and PNHGC, targeted to 23 S rRNA of gram-positive high G+C bacteria and its control probe, we established the most suitable assay conditions. In order to minimize the experiment errors resulted from the differences among stains, we designed another two probes PA-1, PA-2 targeted to 16S rRNA of Actinobacteria based on the alignment of 16S rRNA gene sequence.Using the four probes, PA-1, PA-2, PHGC and PNHGC, we made the identification on the 31 strains of the 56 gram-positive bacteria randomly selected and got 22 positive results, 6 negative results and 3 ambiguous results. In order to confirm the accuracy of FISH result, we detected the...