Anticancer compounds
A technology of compounds and drugs, applied in the field of anticancer compounds
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Embodiment 1
[0204] Example 1: Bacteria Isolation
[0205] The penicillin-producing bacterium Labrenzia PHM005 was isolated in 2005 from a sediment sample collected at a depth of 18 m from a highly epiphytic and unidentified coral sponge habitat off the coast of Kenya. Approximately 5 grams of marine gravel material was collected in 50 ml Falcon tubes containing sterile artificial sea water (ASW) and kept at 5°C for 5 days before processing. Once in the laboratory, samples were homogenized and 100 μl of 1:100 diluted ASW was spread directly onto Petri dishes with 27 g / L sea salt (Tropic PRO-REEF), 16g / L agar and 0.2mg / mL cycloheximide sea salt medium. After three weeks of incubation at 28°C, brownish colonies were picked and transferred to the same sea salt medium to confirm purity and produce biomass for molecular characterization, one colony was inoculated in liquid marine broth , for further storage in 20% glycerol at -80°C as a cell bank.
Embodiment 2
[0206] Example 2: Electron Microscopy.
[0207] Cells in mid-exponential growth phase were adsorbed on 400 mesh carbon-collodion-coated grids for 2 min, negatively stained with 2% uranyl acetate, imaged with a Jeol JEM 1011 transmission electron microscope operated at 100 kV and analyzed with CCDGatan Erlangshen ES1000W camera shooting.
Embodiment 3
[0208] Example 3: 16S rRNA Characterization.
[0209] For DNA extraction, strains were grown in marine liquid medium (DIFCO 1196) for 72 hours. Cells were recovered and lysed by boiling with 4% NP40 for 10 minutes. Cell debris was discarded by centrifugation. The 16S rDNA gene was amplified by polymerase chain reaction using bacterial primers F1 and R5. Using BioNumerics V7.5 (Applied Maths), a phylogenetic tree was generated by performing cluster analysis based on the similarity coefficient and UPGMA of pairwise alignments ( figure 2 ). Phylogenetic neighbors were identified by comparison with the SILVA LTPs123 database, and pairwise 16S rDNA gene sequence similarities were calculated.
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