chapter2Vehicles要点.ppt

* The attraction of M13 as a cloning vehicle –M13作为克隆载体的吸引力 Features of M13 meet the requirements as a cloning vehicle: smaller size, 10kb; The double-stranded replicative form (RF) be easily for experimental purposes. Single-stranded DNA can be obtained from a M13-based vector. –for DNA sequencing and in vitro mutagenesis. * 2.2.3 Viruses as cloning vehicles for other organisms Mammalian viruses: simian virus 40 (SV40): 猿猴病毒40 adenoviruses: 腺病毒 retroviruses: 逆转录病毒 The insect baculoviruses 杆状病毒 当网络经济发展到一定的时候，人们开始关注下一个风险投资的热点，生物技术由此浮出水面。正如比尔·盖茨的预言所说：下一个首富可能是从事生物技术的投资者。 思考题 1 please list essential parts of a cloning vetor，and describe their function 2 classification of plasmid 3 words：plasmid, cccDNA, ori, Episome, copy number, conjugation, compatibility and incompatibility, relaxed and stringent plasmids 当网络经济发展到一定的时候，人们开始关注下一个风险投资的热点，生物技术由此浮出水面。正如比尔·盖茨的预言所说：下一个首富可能是从事生物技术的投资者。 Thanks * Chapter 2 Vectors for Gene Cloning: plasmids and bacteriophages Xinjun Yu, 于欣君 2015.3.17 Contents 2.1 plasmids 2.1.1 Basic features of plasmids 2.1.2 Size and copy number 2.1.3 Conjugation and compatibility 2.1.4. Plasmid classification 2.1.5 Plasmids in organisms other than bacteria The requirements for a DNA molecule as a cloning vectors Self-replication： ori site Special phenotypic traits：genetic marker gene Insert site：Multi-cloning site（MCS） Plasmid size： as small as possible Not affect host cell 2.1.1 Basic features of plasmids Fig.1 Plasmids: independent genetic elements found in bacterial cells Plasmid is an autonomous self-replicating extra-chromosomal circular DNA 特性：1.extra-chromosomal covalently closed circular DNA, cccDNA, size 1-250kbp. 2.Self-replication autonomous replicon. 3. not essential for host. 4. uncompatibility 5. transformation ability Conformations Plasmids are circular molecules of DNA that lead an independent existence in the bacterial cell. Double-stranded circular DNA – covalently c