東方百合‘索邦’組蛋白去乙?；富騆oSorHDA1的克隆與表達分析

LoSorHDA1LoSorHDA1。通過表達分析發(fā)現(xiàn)，LoSorHDA1基因在不同花期和不同組Orientallilyisapreciousflowerwithqualitycharacteristicsmainlydeterminedbygeneregulation.Histonedeacetylasesplayanimportantroleinthecell.Inthisstudy,anewhistonedeacetylasegeneLoSorHDA1wasclonedbyanalyzingandscreeningtheOrientallilygenomedatabase.Throughexpressionanalysis,itwasfoundthatLoSorHDA1genehasdifferentexpressionpatternsindifferentflowerperiodsandtissues,andtheexpressionlevelishighestinflowerbuds.Itisspeculatedthatthisgenemaybeinvolvedintheregulationoffloweringandorgandevelopment.ThisstudyprovidesabasisforfurtherstudyingthegeneregulationmechanismofOrientallily.Keywords:Orientallily,histonedeacetylase,LoSorHDA1,cloning,expressionanalysis,generegulationRNAisoPlus試劑盒、PrimeScriptRTMasterMix、TBGreen?PremixExTaqII試劑盒、TransGenBiotechpEASY-BluntZero載體、pET-28a示例載體和E.coliBL21(DE3)菌株。RNARNAisoPlusRNAcDNAPrimeScriptRTMasterMixRNA樣品進行逆轉(zhuǎn)錄反應生成cDNA。根據(jù)得到的cDNA序列，設(shè)計適當?shù)囊镞M行PCR擴增，將擴增產(chǎn)pEASY-BluntZeroLoSorHDA1基因成功克隆入pET-28a載體并轉(zhuǎn)化至E.coliBL21(DE3)菌株。TBGreen?PremixExTaq?IIqRT-PCR實驗，以8個不同的樣品（包括開花初期、盛花期、衰花期和葉片）為材料考察LoSorHDA1基因的表達模式。LoSorHDA1LoSorHDA14308bpBLAST分析表明，LoSorHDA1基因編碼一種新的組蛋白去乙酰化酶。LoSorHDA1qRT-PCRLoSorHDA1基因在不同花期和不同組生長階段和不同組織中，LoSorHDA1基因的表達量存在很大的差異。在花