基因工程概论重点总结 3-dna重组克隆的单元操作（Introduction to genetic engineering, focusing on the unit operations of 3-DNA recombinant cloning）.docVIP

基因工程概论重点总结 3-dna重组克隆的单元操作（Introduction to genetic engineering, focusing on the unit operations of 3-DNA recombinant cloning） 1. what is the function of the vector used for recombinant DNA cloning? Transporting foreign genes into recipient cells, providing the ability of replication or integration for foreign genes, and providing the necessary conditions for amplification and expression of exogenous genes Four conditions must be met: (1) abundant enzyme sites, which can be transferred to recipient cells, have stable inheritance in recipient cells and have suitable markers for detection. What are the major characteristics of 2. plasmid DNA and virus (bacteriophage) DNA as vectors? Spontaneous replication: independent replication by chromosome DNA Scalability: G- strains, loose replication, and rigorous replication Transferability: binding can be transferred to another cell; genetic engineering uses a non binding type Incompatibility: immunity to infected plasmids or plasmids 3., how to understand the incompatibility of plasmids and its significance in the process of recombinant cloning of DNA Two different plasmids with the same or similar replicon structure or regulatory pattern can not stably exist in the same receptor cell 4. lists the different uses of expressed plasmids, shuttle plasmids, probe plasmids, and COS plasmids Gao Kaobei: a mutant copy number, 1000-3000 copy, used to amplify genes Low copy: from pSC101, copy less than 10, expressing toxic genes Wen Min: different copies are displayed at different temperatures Sequencing: contains sequencing, universal primers, complementary sequences and multiple enzyme connectors Integration: incorporating integration promoting genes and loci that facilitate the integration of foreign genes onto chromosome DNA Shuttle: contains two different copies of the receptor for cloning, easy for gene cloning Expression: transcription, translation, and purification of a gene with enhanced foreign gene expression Probe: equipped