复旦大学生物化学课件-DNA的生物合成和损伤修复.pptVIP

Section One The General Features of Replication of Chromosomal DNA DNA replicates semiconservatively. ⑴. Watson and Crick predicted that DNA might semiconservatively replicate. the hypothesis ___________ p ___________ d ___________ p ___________ p ____________ d ____________ p ⑵. In 1958 Meselson and Stahl demons- trated the semiconservative nature of DNA replication in E coli. The experiment: CsCl equilibrium gradient density ultracentri- fugation of 15N labeled E coli DNA. The density of DNA was increased by labeling it with 15N, a heavy isotope of nitrogen. This was done by growing E coli 15 ge- nerations in a medium that contained 15NH4Cl as its only nitrogen source. 15N DNA was extracted and subjected to CsCl equilibrium gradient density ultracen- trifugation. The DNA band position was recorded. There was one band of 15N DNA. The bacteria were transferred to an 14NH4Cl medium and grown for one generation. The DNA density was determined again. The position of the DNA band was compared with that of the 15N DNA . What was the result ? There was one DNA band. It had a lower density than 15N DNA because its position was above on that of 15N DNA. It was 15N/14N hybrid DNA. After another generation growing in the 14NH4Cl medium the bacterial DNA density was determined. There were two DNA bands. One half of the DNA was 14N DNA, and another half was hybrid DNA. In succeeding generations the ratio of 14N DNA to hybrid DNA increased gradually. The hybrid DNA became less and less.. summary DNA replicates in a semiconservative man- ner. When the two parental strands sepa- rate, each serves as the template for