预览加载中,请您耐心等待几秒...
1/10
2/10
3/10
4/10
5/10
6/10
7/10
8/10
9/10
10/10

亲,该文档总共33页,到这已经超出免费预览范围,如果喜欢就直接下载吧~

如果您无法下载资料,请参考说明:

1、部分资料下载需要金币,请确保您的账户上有足够的金币

2、已购买过的文档,再次下载不重复扣费

3、资料包下载后请先用软件解压,在使用对应软件打开

主要内容一、转基因(Transgenesis)技术1、微生物中的基因转移2、植物转基因技术3、动物转基因技术二、基因敲除技术1.利用基因同源重组进行基因敲除2.利用随机插入突变进行基因敲除 ——基因捕获法3.RNA干扰引起的基因敲除基因敲除技术与诺贝尔奖文献: Constructionofpha-Operon-DefinedKnockoutMutantsofPseudomonas putidaKT2442andtheirApplicationsinPoly(hydroxyalkanoate)Production PseudomonasputidaKT2442couldaccumulatemedium-chain-lengthpoly(hydroxyalkanoate)s(PHA)consistingof3-hydroxyhexanoate,3-hydroxyoctanoate,3-hydroxydecanoate,and3-hydroxydodecanoatefromawiderangeofcarbonsources.Inthisstudy,thePHAsynthasephaoperon(phaC1-phaZ-phaC2)wasknockedoutandthevgbgeneencodingvitreoscillahemoglobinprotein(VHb),whichcouldenhanceoxygenuptakerateespeciallyatlowoxygenconcentration,wasintegratedintotheP.putidaKT2442genometoreplacethedeletedfragment.TheresultingmutantP.putidaKTOY01orgene-replacedmutantKTOY02wasusedasthehosttostudyPHAsynthasepropertiesandPHAproduction.DifferentPHApolymerase(PhaC)genes,phaCRefromRastoniaeutrophaH16,phaCAcfromAeromonascavie,andphaC2PsfromPseudomonasstutzeri1317,wereexpressedinthemutantstrainstotestthePhaCenzymesubstratespecificity.TheresultshowedP.putidaKTOY01orKTOY02couldprovidenotonlymclPHAmonomersbutalso3-hydroxybutyratefromfattyacids,whichmayallowtheproductionofcopolyesterspoly(3HB-co-mcl3HA).PlasmidpCJY10containingphaC2Ps,phbA,andphbBgenesencodingPHApolymerase,b-ketothiolase,andacetoacetyl-CoAreductase,respectively,weretransformedintoP.putidaKTOY01andKTOY02.Shake-flaskcultureshowedP.putidaKTOY01orKTOY02(pCJY10)couldaccumulatepoly(3HB-co-mcl3HA)fromglucose.TheaboveresultshowedphaoperonknockoutmutantofP.putidaKT2442wasaveryusefulhostofgreatpotentialnotonlyforstudyingPhaCsynthase,butalsoformicrobialproductionofcopolyesterspoly(3HB-co-mcl3HA),whichisverydifficulttoabtain.RECOMBINATIONaA1A1致谢补充资料