原生质体分离试验方法ppt

Ahighlyefficientricegreentissueprotoplastsystemfortransientgeneexpressionandstudyinglight/chloroplast-relatedprocesses水稻原生质体分离与GFP，BiFC等方法研究意义植物原生质体是常见的与生理相关的细胞组织，可用于分析基因的功能特性，有重要意义。在水稻研究中，有关悬浮培养细胞和黄化苗原生质体研究较多，但绿色组织原生质体研究较少。绿色植物原生质体成功用于植物与激素、代谢物及环境因素相关的信号转导的研究。简化并提高了基因短暂表达的系统，并结合蛋白质免疫印染、点位及蛋白质-蛋白质相关试验。主要内容Here,wereportasimplifiedmethodforisolatingprotoplastsfromnormallycultivatedyoungricegreentissuewithouttheneedforunnecessarychemicalsandavacuumdevice.主要内容Transfectionsofthegeneratedprotoplastswithplasmidsofawiderangeofsizes(4.5-13kb)andco-transfectionswithmultipleplasmidsachievedimpressivelyhighefficienciesandallowedevaluationsby1)proteinimmunoblottinganalysis,2)subcellularlocalizationassays,3)protein-proteininteractionanalysis.图片GFPChlMergedBrightGFPChlMergedBright问题7页：Theupperpanelcontainslowpowerimagesshowinghighco-expressionefficiencyperoxisomes过氧化物酶体研究结果Importantly,thericegreentissueprotoplastswerephotosyntheticallyactiveandsensitivetotheretrogradeplastidsignalinginducernorflurazon(NF).TransientexpressionoftheGFP-taggedlight-relatedtranscriptionfactorOsGLK1markedlyupregulatedtranscriptlevelsoftheendogeneousphotosyntheticgenesOsLhcb1,OsLhcp,GADPHandRbcS,whichwerereducedtosomeextentbyNFtreatmentinthericegreentissueprotoplasts.