园艺植物生物技术第七章.pptxVIP

园艺植物生物技术第七章;Enhancer element;Genetic Engineering also called genetic modificat is the direct manipulation of an organisms genome using biotechnology. New DNA may be inserted in the host genome by first isolating and copying the genetic material of interest using molecular cloning methods to generate a DNA sequence, or by synthesizing the DNA, and then inserting this construct into host organism. Genes may be removed, or knocked out, using a nuclease. Gene targeting is a different technique that uses homologous recombination to change an endogenous gene, and can be used to delete a gene, remove exons, add a gene, or introduce point mutations; An organism that is generated through geneti engineering is considered to be a geneticall modified organism (GMO). The first GMOs were bacteria in 1973. Genetically modified crops (GMCs, GM crops).The first genetically modified plant produced in 1982, using an antibiotic-resist tobacco plant. The first genetically modifie crop approved for sale in the U.S., in 1994, w the FlavrSavr tomato, which had a longer shel life.;7.1 Instrumental Enzyme of Gene Engineering;7.1.1 Restriction Endonuclease;TypeⅡendonuclease;Isoschizomers(同裂酶)：指来源不同，但识别相同靶序列的核酸内切酶。 如：HpaⅡ和MspⅠ是一对同裂酶(CCGG) Isocaudarner(同尾酶)：指来源不同、识别靶序列不同但产生相同的粘性末端的核酸内切酶。;7.1.2 DNA ligase （DNA连接酶）;7.1.3 DNA polymerase;Other DNA-modifying enzymes;7.2 Vectors（载体）;7.2.1 Plasmids(质粒);（4363bp）;pUC Vectors;第十六页，共九十八页，2022年，8月28日;7.2.2 λ phage(噬菌体) vector;Vector carry LacZ gene：;7.2.3 Cosmid(柯斯质粒) vectors;COSmid;7.2.4 Artificial chromosome vector;Structure of a bacterial artificial chromosome (BAC), used for cloning large fragme of donor DNA. CMR is a selectable marker for chloramphenicol resistance. or repE, parA, and parB are F genes for replication and regulation of copy number. cosN is the cos site from l phage. HindIII and BamHI are cloning sites at which foreign DNA is inserted. The two promoters are for transcribing the inserted fragment. The NotI sites are used for cutting out