Expression of Pyruvate Carboxylase mRNA Variants in Liver of Dairy Cattle at Calving 英文参考文献.docVIP

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Expression of Pyruvate Carboxylase mRNA Variants in Liver of Dairy Cattle at Calving 英文参考文献.doc

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Expression of Pyruvate Carboxylase mRNA Variants in Liver of Dairy Cattle at Calving 英文参考文献

ExpressionofPyruvateCarboxylasemRNAVariantsin LiverofDairyCattleatCalving CansuAgca¤,ShawnS.Donkin* DepartmentofAnimalSciences,PurdueUniversity,WestLafayette,Indiana,UnitedStatesofAmerica Background. Bovine liver expresses six pyruvate carboxylase (PC) transcript variants, bPC59A, bPC59B, bPC59C, bPC59D, bPC59E,andbPC59F,whichonlydifferatthe59untranslatedregion(UTR)andcontainacommoncodingregion.Theobjective of this experiment wasto determine the profile and abundance of PC transcripts in bovine liver at calving. Methodology/ PrincipalFindings.Aribonucleaseprotectionassay(RPA)protocolwasdevelopedtosimplifyanalysisofthesevariantsand investigatethechangesinabundanceofeach59UTRtranscriptrelativetototalPCmRNA.Liverbiopsysamplescollectedfrom sevencowson+1drelativetocalvingwereanalyzedbyRPAtodeterminetheprofileinPC59UTRvariants.Resultsshowthat allsixbovinePC59UTRvariantsaredetectedatcalving.DataindicatethatbovinePC59UTRvariantAisthemostabundant, variants C and E are least abundant and expression of variants B, D and F is intermediate at calving. Conclusions. This manuscriptdescribesasimplifiedRPAmethodthatquantifiestheabundanceofsixPCtranscriptsbyusingtworiboprobes. ThelackofuniformityinthepatternofPC59UTRvariantsatcalvingsuggestsanadditionalcomplexityforcontrolofbovinePC mRNAexpressionatcalvingthatmaybetheresultoftranscriptionalcontrols,variationinmRNAprocessing,oracombination oftheseprocesses. Citation: AgcaC,DonkinSS(2007)ExpressionofPyruvateCarboxylasemRNAVariantsinLiverofDairyCattleatCalving.PLoSONE2(12):e1270. doi:10.1371/journal.pone.0001270 INTRODUCTION plasmids contain 90bp of the coding region in addition to their respective59UTRs.ThetruncatedplasmidsABCEFandBCDE Pyruvate carboxylase plays an essential role in many metabolic pathways including gluconeogenesis, lipogenesis, amino acid metabolism and neurotransmitter synthesis. PC is a flux- generatingenzymeforseveralmetabolicpathwaysanditsactivity iscontrolledbyallostericactivationbyacetyl-CoA,transcriptio