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Transformation and transfection Packaging phage DNA in vitro Alternative DNA delivery methods 本章要點(diǎn): 掌握基因工程受提細(xì)胞的類型和要求。 掌握載體的基本結(jié)構(gòu)和質(zhì)粒載體、噬菌體載體的特點(diǎn)、構(gòu)建原理。 掌握在體外源 DNA 連接的方式和原理 以及感受態(tài)菌的制備原理。 掌握基因轉(zhuǎn)入受體細(xì)胞的基本方法和原理。(重點(diǎn)) Chapter 6 Cloning strategies (6hrs) which approach is best Cloning from mRNA Synthesis of cDNA Cloning cDNA in plasmid vectors Cloning cDNA in bacteriophage vectors Cloning from genomic DNA Genomic libraries Preparation of DNA fragments for cloning Ligation, packaging and amplification libraries Advanced cloning strategies Synthesis and cloning of cDNA Expression of cloned cDNA molecules Cloning large DNA fragments BAC and YAC vectors 本章要點(diǎn):掌握化學(xué)合成 DNA的原理和 DNA 文庫的構(gòu)建原理。 了解酵母人 工染色體和細(xì)菌人工染色體的結(jié)構(gòu)和工作原理。 ( 次 重點(diǎn)) Chapter 7 The polymerase Chain reaction (4hrs) The history of PCR The methodology of the PCR The essential features of the PCR The design of primers for PCR DNA polymerases for PCR More exotic PCR techniques PCR using mRNA templates Nested PCR Inverse PCR RAPD and several other acronyms Processing of PCR products Application of the PCR 本章要點(diǎn):掌握 PCR 擴(kuò)增 DNA 的技術(shù)原理和方法。(重點(diǎn)) Chapter 8 Selection, Screening and analysis of rebinants (6hrs) Geic selection and screening methods The use of chromogenic substrates Insertional inactivation Complementation of defined mutations Other geic selection methods Screening using nucleic acid hybridisation Nucleic acid and probes Screening clone banks Immunological screening for expressed genes Analysis of cloned genes Characterisation based on mRNA translation in vitro Restriction mappin Blotting techniques DNA sequencing 本章要點(diǎn):掌握常用的抗菌素抗性篩選、酶切篩選、 雜交篩選、 酶聯(lián)免疫吸附測定( ELISA)、免疫印跡( western blotting)法的原理。了解轉(zhuǎn)譯篩選法和真核生物的報(bào)道基因篩選法。 (重點(diǎn)) Part III Geic Engineering in Action (12hrs) Chapter 9 Understanding genes and genomes (self study) Analysis of gene structure and function A closer look at sequences Finding important