Roseobacter denitrificans OCh 114

Roseobacter denitrificans OCh 114.A single colony of Roseobacter denitrificans strain OCh 114 was grown heterotrophically and total DNA was isolated using proteinase K treatment followed by phenol extraction. The DNA was fragmented by kinetic shearing, and three shotgun libraries were generated: small and medium insert libraries in pOTWI3 (using size fractions of 2-3 kb and 6-8 kb, respectively), and a large insert fosmid library in pEpiFOS-5 (insert sizes ranging from 28-47 kb), which was used as a scaffold. The relative amount of sequence coverage obtained from the small, medium, and large insert libraries was 8X, 1X, and 1X, respectively. The whole genome sequence was established from 55,081 end sequences (giving 9.6X coverage) derived from these libraries using dye terminator chemistry on ABI 3730xl automated sequencers.The aerobic phototrophic bacteria are ubiquitous as plant symbionts, free-living in lakes and ocean surface waters, soils and even near deep sea hydrothermal vents. In fact, ~10% of all microorganisms in marine surface water samples appear to be aerobic phototrophic bacteria, suggesting a significant contribution to the global carbon cycle. Although the numbers of aerobic phototrophic bacteria vary with locale (Goericke, 2002), it is clear that these organisms are a previously unrecognized major component of the bacterial biomass on Earth. The genome sequence of this representative aerobic phototrophic bacteria species will enable discovery in the following three areas: 1) the evolutionary genesis of photosynthesis genes; 2) pathways of carbon dioxide fixation and production; 3) light and oxygen signal transduction in gene expression.Goericke R (2002) Bacteriochlorophyll a in the ocean: Is anoxygenic bacterial photosynthesis important? Limnol Oceanogr 47: 290-295. (NCBI BioProject: bp_list[1])