C3： DNA replication - 修 分子生物学双语课程PPT.pptVIP

Chapter 03: DNA Replication 3.1 The principle of DNA replication 3.2 DNA replication model 3.3 Enzymes and protein needed in DNA replication 3.4 Process of DNA replication 3.5 Telomere and Telomerase 3.1 The principle of DNA replication 3.1.1 semi-conservative replication Definition: during replication, the two parental strands separate and each acts as a template to direct the synthesis of a new complementary daughter strand. DNA replication pattern: conservative model、semi-conservative model、dispersive model 1968 Okazaki radioactive label (3H-dTTP) + CsCl density gradient centrifugation → label enters newly synthesized DNA in the form of short fragments(~1000-2000 bases) The ligase temperature-sensitive mutants → under the temperature in which the ligase is inactive → a large amount of short fragments but no lagging strand Conclusion: The lagging strand must be synthesized in the form of Okazaki fragments On the leading strand, DNA synthesis can proceed continuously in the 5’ to 3’ direction as the parental duplex is unwound. On the lagging strand, a stretch of single-stranded parental DNA must be exposed, and then a segment is synthesized in the reverse direction (relative to fork movement). A series of these fragments are synthesized, then they are joined together to create an intact lagging strand. 3.1.3 Primer A common feature of all DNA polymerases is that they cannot initiate synthesis of a DNA chain de novo. All DNA polymerases require a 3’-OH end to initiate DNA synthesis. For DNA replication, a special RNA polymerase called a primase synthesizes an RNA chain(primer) that provides the priming end. Purpose: keep the high faithfulness Reason: DNA polymerase has proofreading activity, while RNA polymerase dont. After the low-fidelity primer complete its function, it will be replaced by high-fidelity DNA synthesized DNA polymerase. Results: improve the accuracy of the DNA replication up to 105 times. 3.2 Replication Model Replicon：means a s