第七章园艺植物生物技术GeneticEngineeringalsocalledgeneticmodification,isthedirectmanipulationofanorganism'sgenomeusingbiotechnology、NewDNAmaybeinsertedinthehostgenomebyfirstisolatingandcopyingthegeneticmaterialofinterestusingmolecularcloningmethodstogenerateaDNAsequence,orbysynthesizingtheDNA,andtheninsertingthisconstructintothehostorganism、Genesmayberemoved,or"knockedout",usinganuclease、Genetargetingisadifferenttechniquethatuseshomologousrebinationtochangeanendogenousgene,andcanbeusedtodeleteagene,removeexons,addagene,orintroducepointmutations、Anorganismthatisgeneratedthroughgeneticengineeringisconsideredtobeageneticallymodifiedorganism(GMO)、ThefirstGMOswerebacteriain1973、Geneticallymodifiedcrops(GMCs,GMcrops)、Thefirstgeneticallymodifiedplantwasproducedin1982,usinganantibiotic-resistanttobaccoplant、ThefirstgeneticallymodifiedcropapprovedforsaleintheU、S、,in1994,wastheFlavrSavrtomato,whichhadalongershelflife、7、1InstrumentalEnzymeofGeneEngineering7、1、1RestrictionEndonucleaseTypeⅡendonucleaseIsoschizomers(同裂酶):指来源不同,但识别相同靶序列得核酸内切酶。如:HpaⅡ与MspⅠ就是一对同裂酶(CCGG)Isocaudarner(同尾酶):指来源不同、识别靶序列不同但产生相同得粘性末端得核酸内切酶。BamHⅠBclⅠGGATCCTGATCACCTAGGACTAGT7、1、2DNAligase(DNA连接酶)7、1、3DNApolymerase7、1、4OtherDNA-modifyingenzymes7、2Vectors(载体)127、2、1Plasmids(质粒)(4363bp)pUCVectors7、2、2λphage(噬菌体)vector7、2、3Cosmid(柯斯质粒)vectorsCOSmidpHC79is4、3kbDNAfragmentandmadeofpBR322andcossiteofλphage、InsectionDNAfragmentmorethan40kb。7、2、4ArtificialchromosomevectorsStructureofabacterialartificialchromosome(BAC),usedforcloninglargefragmentsofdonorDNA、CMRisaselectablemarkerforchloramphenicolresistance、oriS,repE,parA,andparBareFgenesforreplicationandregulationofcopynumber、cosNisthecossitefromlphage、HindIIIandBamHIarecloningsitesatwhichforeignDNAisinserted、Thetwopromotersarefortranscribingtheinsertedfragment、TheNotIsitesareusedforcuttingouttheinsertedfragment、7、3GenecloningGenelibraryscreening(文库筛选法)Map-basedcloning(图位克隆技术)Transposontag(转座子标签法)Differentialdisplayanalysis(基因差异表达分析)Homologysequences(同源序列法)Genechip(