[自然科学]上海交大生物化学课件Chapter6.ppt

Chapter 6  Exploring Genes Recombinant DNA technology, a new approach to exploring the central molecules of life A powerful means of analyzing and altering genes and proteins. as a fruit of several decades of basic research on DNA, RNA and viruses. It depends on having enzymes that can cut, join, and replicate DNA and reverse-transcribe RNA (restriction enzymes, DNA ligases连接酶, DNA polymerases, reverse transcriptase,). Thus, it is based on nucleic acid enzymology. DNA重组技术一靠核酸酶学,二靠碱基配对语言 A second foundation is the base-pairing language that mediates nucleic acid recognition. hybridization with complementary DNA or RNA probes is a sensitive and powerful means of detecting specific nucleotide sequence. 与探针杂交 In recombinant DNA technology, base pairing is used to construct new combinations of DNA as well as to detect and amplify particular sequences. This revolutionary technology is also critically dependent on the existence of viruses and plasmids, 三靠载体(病毒和质粒) some of the benefits of these new methods the discovery of restriction enzymes限制酶 led to the development of techniques for the rapid sequencing of DNA. A wealth of information concerning gene architecture, the control of gene expression, and protein structure has come from the sequencing DNA molecules.(限制酶 →测序→各种信息) DNA molecules can also be synthesized de novo. The automated solid-phase synthesis of DNA provides highly specific probes and synthetic tailor-made genes. 提供高度特异的探针和合成量身定做的基因 A single copy of a specific DNA sequence in a complex mixture can be amplified more than a millionfold by the polymerase chain reaction PCR. the construction and cloning of novel combinations of genes New genes can be efficiently expressed by host cells, as exemplified by the production of human insulin by bacteria. Moreover, specific mutations can be made in vitro to engineer proteins in designed ways. 基因重组→有效表达→(经定向突变)→工程制造特定蛋白 Restriction Enzymes Split DNA into Specific Fragments Restriction enzymes, also