分子生物学全套课件96P课件.pptVIP

Serial analysis of gene expression, SAGE 34 million base pair nuclear genome 129 thousand base pair plastid genome 44 thousand base pair mitochondrial genome 24 diploid nuclear chromosomes novel genes for silicic acid transport and formation of silica-based cell walls high-affinity iron uptake biosynthetic enzymes for several types of polyunsaturated fatty acids use of a range of nitrogenous compounds a complete urea cycle All attributes that allow diatoms to prosper in aquatic environments more? Draft nuclear genome of the marine diatom Thalassiosira pseudonana DNA isolation Genomic DNA was isolated using Hoechst dye and cesium chloride centrifugation to separate nuclear from mitochondrial and plastid DNA A single cell isolate was originally collected from Moriches Bay, Long Island Sound, New York in 1958, and has been maintained continuously in culture since then as CCMP 1335 (Provasoli-Guillard National Center for Culture of Marine Phytoplankton; /) This cell line has at least twice been reduced to a single cell, once during its maintenance and again in preparation for DNA purification for genome Sequencing (cloning, pure culture) Asexually propagated (mitotic) pure population Life history of Ditylum brightwellii (diatom) Preparative density-gradient ultracentrifugation of DNA Under high centrifugal force, a solution of cesium chloride (CsCl) molecules will dissociate, and the heavy Cs+ atoms will be forced? towards the outer end of the tube, thus forming a shallow density gradient. DNA molecules placed in this gradient will migrate to the point where they have the same density as the gradient (the neutral buoyancy or isopycnic point). The gradient is sufficient to separate types of DNA with slight differences in density due to differing [G+C] content, or physical form (e.g., linear versus circular molecules). ? After centrifugation for 10 hrs at 100,000 rpm (450,000 x g), two distinct bands, corresponding to sheared linear nuclear DNA above and