i need fulldetails about isolation and characeristics of streptomyces...?

Question

test for streptomyces with images

Answer 1

Upstream of the Streptomyces coelicolor A3(2) chitinase G gene, a small gene (named chb3) is located whose deduced product shares 37% identical amino acids with the previously described CHB1 protein from Streptomyces olivaceoviridis. The chb3 gene and its upstream region were cloned in a multicopy vector and transformed into the plasmid-free Streptomyces lividans TK21 strain. The CHB3 protein (14.9 kDa) was secreted by the S. lividans TK21 transformant during growth in the presence of glucose, N-acetylglucosamine, yeast extract, and chitin. The protein was purified to homogeneity using anionic exchange, hydrophobic interaction chromatographies, and gel filtration. In contrast to CHB1, CHB3 targets α-chitin, β-chitin, and chitosan at pH 6.0 but does so relatively loosely. The ecological implications of the divergence of substrate specificity of various types of chitin-binding proteins are described.

Streptomyces spp. are soil bacteria and main decomposers of chitin, the second most abundant polysaccharide in nature, in soil. Since streptomycetes use chitin as carbon and nitrogen sources, they are assumed to play a major part in the turnover of chitin in natural ecosystems. A number of chitinases have been characterized from various Streptomyces spp. (1, 2, 7, 10, 20, 27, 29). Several genes encoding chitinases (EC 3.2.1.14) that hydrolyze chitin have been cloned from Streptomyces spp. (2, 3, 6, 15, 16, 19–21, 28). The deduced proteins belong to family 18 or 19 of glycosyl hydrolases (3, 19).

The chitin-binding proteins CHB1 and CHB2 lacking hydrolytic activity have been found in Streptomyces olivaceoviridis and Streptomyces reticuli, respectively (11, 25). These proteins specifically bind to α-chitin but not to β-chitin, chitosan, or cellulose. The CBP21 protein produced by a gram-negative, chitin-degrading bacterium, Serratia marcescens (26), shares 45% identical amino acids with CHB1.

We previously found one open reading frame upstream of the Streptomyces coelicolor A3(2) chitinase gene chiG. Its deduced product (21) shares 37% identical amino acids with the previously described chitin-binding protein CHB1 (25). In this report we describe the biochemical characteristics of the protein and give some insights into the regulation of the corresponding gene.

Bacterial strains, plasmids, and selection of transformants. Escherichia coli DH5α (23) was used as host for constructs derived from the vector pUC18 (31) or the bifunctional Escherichia coli-Streptomyces vector pWHM3 (30). E. coli DH5α transformants carrying pUC18 or pWHM3 constructs were selected on Luria-Bertani medium agar plates containing ampicillin (50 μg/ml).

TK21, a plasmid-less derivative from Streptomyces lividans 66 (9), served as the host for the pWHM3-based construct (named pWF17) carrying the chb3 gene. Protoplasts from S. lividans TK21 were transformed as described elsewhere (9). Regenerated transformants containing pWHM3-based constructs were selected on complete medium containing thiostrepton (25 μg/ml).

Recombinant DNA techniques and transformations. Digestions by restriction enzymes were performed as described earlier (23) and according to the suppliers' instructions. Dephosphorylation, blunting, or ligation of DNA fragments was done using bacterial alkaline phosphatase, a DNA blunting kit, or a DNA ligation kit, respectively, according to the manufacturers' instructions. Transformations of E. coli and S. lividans were performed as described previously (9, 23). Bacterial alkaline phosphatase for DNA manipulation was bought from Toyobo, Tokyo, Japan. A DNA blunting kit and a DNA ligation kit were obtained from Takara, Tokyo, Japan. Restriction enzymes were supplied by Gibco-BRL and Roche Diagnostics.

Answer 2

Physical morphology: Long chains of filimentous rods
Aerobic
Gram Stain: Not acid fast
usually cause the disease entity known as mycetoma (fungus tumor).
Material sent to the lab is pus or skin biopsy
can grow on both bacterial and fungal (Sabouraud) media
actinomycetes take longer to be visible on the culture plates.
Rough dry colonies
Catalase Positive
Oxidase Negative
(Some strains produce antibiotics) Imbed a colony of strep. over E. Coli and look for zone of inhibition

Answer 3

Physical morphology: Long chains of filimentous rods
Aerobic
Gram Stain: Not acid fast
usually cause the disease entity known as mycetoma (fungus tumor).

Answer 4

See this website.
http://streptomyces.org.uk/