增殖细胞核抗原基因作为东海原甲藻生长指标的现场实验分析

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1、(3)将PCNA相对表达量法与现场培养法所得的生长率作比较，发现二肴具有很好的一致性，都能反映现场水体中赤潮生物的原位生长情况。将该方法应用到2011年春季长江口外和舟山外海赤潮发生区的东海原甲藻生长率现场测定，发现多数站位生长率为负值，表明此次赤潮已经进入消亡阶段。本研究将PCNA相对表达量法应用于现场，对东海原甲藻赤潮进行生长率测定，很好地反映了赤潮的生长状况和发展趋势，为运用分子生物学技术对赤潮进行监控和预报奠定了基础。关键词：赤潮；东海原甲藻：生长率：相对表达量：增殖细胞核抗原Thestudyoftheproliferatingcellnuclearantigengeneasth

2、egrowthindicatorofProrocentrumdonghaienseLuinthefieldstudyAbstractmensurationofthephysiologicalandecologicalparameterssuchastheinsitugrowthrateisthebasicofunderstandingtheregulationofphytoplanktondynamics，constructingtheecologicaldynamicmodelsandevenforecastingthehappeningofredtideInthepast，there

3、searchinourlabshowsthat，therelativeexpressionleavelofproliferatingcellnuclearantigen(PCNA、genecanbeusedasagoodindicatortothephytoplanktongrowthrate，moreover，establishedaformulabetweentherelativeexpressionlevelofPCNAgeneandthegrowthratep(d“)aboutSkeletonemacostatumandProrocentrumdonghaienseItpaves

4、awayformonitorandpredictingtheredtideInthisstudy，basedonthepreliminaryresearchresults，wepayaattentiontousingthismethodstomeasuretheinsitugrow',hrateofthefiledsamplesWedosomeresearchonthesefieldssuch拈thecollectionmethods，themethodsofexlractingtheRNA，improvingthespecificityofthepalmersabouttherefer

5、encegeneandoptimizingthereactionsystemofRFO—PCRinordertodeterminetheinsitugrowthrateofthesamplesfromtheProrocentrumdonghaienseredtideareaintheEastChinaSea，weusetheestablishedmethodthatmeasuretherelativeexpressionlevelofPCNAgenetogetgrowlhrote，Themethodw∞validatedandapplied，enumeratingtheredtidemo

6、nitoraccuratelyusingthismethodsmainresultsRfe弘foIlows：(1)WeresearehofftheefficiencyofRNAextractionincludingthenumberofcells，grindingmethods，membranespecificationsandextractionreagentsTodeterminethelowestnumberofsamplesis2x106cellsTococllectthesamplesbyfilteringthewaterwithcelluloseacetatemembrane

7、whichdiameteris50mm．poreis10gmDealingwiththesamples，Callgrindthemembranewithliquidnitrogen，orgfindthemicroalgaescrapedo仃withliquidnitrogen．dependingonsamplesInextractionreagent，wechooseTRIZOLonextractionfirstly(2)Thepd