分子生物学chapter6-The-Structure-of-DNA-and-RNA.pptVIP

CHAPTER 6 The Structures of DNA and RNA DNA STRUCTURE (1) denaturation of DNA: The process of strand separation is called denaturation Denaturation can be caused by high temperature, high pH Hyperchromicity: when DNA is denatured, the optical density at 260 nm markedly increases. Absorbance is also used to check the DNA or RNA purity A260/A280: dsDNA--1.8 pure RNA--2.0 protein--0.5 The Tm is the midpoint of the temperature range for denaturation. Tm is largely determined by G:C content and ionic strength Annealing Annealing of DNA describes the renaturation of a duplex structure from single strands that were obtained by denaturing duplex DNA. Complementary single strands can renature when the temperature is reduced. hybridization Renaturation / Annealing can occur with DNA-DNA,DNA-RNA, or RNA-RNA combinations, and can be intermolecular or intramolecular. DNA TOPOLOGY DNA TOPOLOGY (1) Structure (1): Linking number is an invariant topological property of covalently closed, circular DNA (cccDNA) Linking number（连环数）is the number of times one strand have to be passed through the other strand in order for the two strands to be entirely separated from each other. Species of cccDNA Plasmid and circular bacterial chromosomes Linear DNA molecules of eukaryotic chromosomes due to their extreme length, entrainment in chromatin and interaction with other cellular components (Ch 7) Structure (2): Linking number is composed of Twist and Writhe The linking number is the sum of the twist （扭转数）and the writhe（缠绕数）. Twist is the number of times one strand completely wraps around the other strand. Writhe is the number of times that the long axis of the double helical DNA crosses over itself in 3-D space. DNA TOPOLOGY (2) Local disruption of base pairs Function (1): DNA in cells is negatively supercoiled; nucleosomes introduces negative supercoiling in eukaryotes Negative supercoils serve as a store of free energy that aids