Molecular characterization and expression profiles of Sp-Ub during gonad development in Scylla paramamosain 
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1. Key Laboratory of Healthy Mariculture in the East China Sea, Ministry of Agriculture, Fisheries College, Jimei University, Xiamen, 361021, China;

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S968

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    Abstract:

    isaneconomicallyimportantbreedingcrabfoundalongthesoutheastcoastofChina.Althoughextensiveresearchonreproductivephysiologyhasbeencarriedoutforthisspecies,descriptionoftheregulatorymechanismofthegonaddevelopmentatamolecularlevelislimited.Withinalltissues,themajorityofintracellularproteinsaredegradedbytheubiquitin(Ub)–proteasomepathway(UPP).Inthispathway,UbisconjugatedtoproteinsthataredestinedfordegradationbyanATP-dependentprocessthatinvolvesthreeenzymes.Todate,somestudieshavediscoveredthatcomponentsoftheubiquitinsystemmaybeinvolvedinvariousstepsandprocessesduringgametogenesis,modulationofsteroidreceptorconcentrations,placentaldevelopmentandendometrialmodificationatthebeginningofpregnancy.However,researchontheactionofUPPingonaddevelopmentincrabspeciesUbgene()wasidentifiedandcharacterizedusinganapproachwhichcombinestheexpressedsequencetag(EST)methodandrapidamplificationofcDNAend(RACE).Theexpressionprofileofinthevarioustissuesanddifferentdevelopingstagesofthegonadwasdeterminedbyreal-timequantitativePCR.ThefulllengthcDNAofthegeneisof555bp.Itencodesaproteinof154aminoacids,inwhichthe1-76aminoacidsfallwithintheubiquitindomain,andthe101147aminoacidsfallwithintheribosomal_S27adomain.ThesecondstructurepredictionrevealedthattheubiquitindomainoftheSp-Ubproteincontainsfiveextendedstrands,anα-helixandsomerandomcoils.Moreover,thethree-dimensionalstructurepredictedbytheSWISS-MODELshowedthatitcontainsanα-helixandfourβ-pleatedsheets.AlignmentoftheaminoacidsequenceswithindifferentUbproteinsindicatedthattheubiquitindomainandtheribosomal_S27adomainamongdifferentspeciesarehighlyconservedduring.Real-timequantitativePCRresultsshowedthatwasexpressedbytheubiquitindomaininalltissuesexamined(P>0.05)whichillustratedthattheUbgeneplaysawidevarietyofrolesinbiologicalprocesses.TheexpressionprofilesofinthedifferentdevelopingstagesoftheovaryrevealedthatitsexpressionlevelwasthehighestatstageO5followingstageO1,O4andO3.ItwasthelowestlevelatstageO2whichwassignificantlylowerthanstageO5(P<0.05).Withinthedevelopingstagesofthetestis,theexpressionlevelofwashigherinstageT2followingstageT1andT3,anditsexpressionlevelwassignificantlyhigherinstageT2comparedtostageT3(P<0.05).Duringgametogenesis,theoogoniaandspermatogoniumundergoactivemitosisproliferationforincreasingtheirnumberattheproliferationstage.Previousresearchshowedthatcyclin/Cdksworkcloselywiththeubiquitin-proteasomepathwayduringthemitoticcycle.Thus,itisreasonabletoobservethattheexpressionlevelofexpressionlevelatstageO2maybeduetotheinitiationofformationofprimaryoocytes,andinitialvitellogenesiswithalowlevelofproliferation.Moreover,theexpressionlevelofgraduallyincreasedastheyolkproteinswereaccumulatinginthegrowingoocytesduringvitellogenesis.Amassofproteinsneededtobesynthesizedanddegradedduringgametogenesis,andtheevidenceofubiquitin–conjugatedvitellinpolypeptidesininsectoocyteshadbeenreported.Theseresultsindicatedthatmaybeinvolvedinvitellogenesis.Furthermore,thetestisofcrabcontainsgermcellsofvariousdevelopmentstageswithintheseminiferoustubules.Hence,thehigherexpressionlevelofdevelopmentmaybenecessaryforfastincreasingthenumberofspermatocytecells,whichindicatedthatUPPwasmoreactiveinthesecells.Conclusively,ourstudiesarethefirstreportofcloningandexpressionprofilingofUbincrustaceans.AlloftheseresultscanprovidebasicdatashowingtheroleofSp-Ubwithinthegonaddevelopmentofthecrab.

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戴燕彬,韩坤煌,颜素芬,邹志华,张子平,王艺磊. 拟穴青蟹泛素基因的克隆及其在性腺发育过程中的表达[J]. Jounal of Fishery Sciences of China, 2012,[volume_no](6):946-955

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  • Online: November 08,2012
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