马鹿cervuselaphus鹿茸角化细胞生长因子kgf:基因克隆、序列分析及转录模式研究-cervuselaphus antler keratinocyte growth factor kgf gene cloning, sequence analysis and transcription pattern research.docxVIP

马鹿cervuselaphus鹿茸角化细胞生长因子kgf:基因克隆、序列分析及转录模式研究-cervuselaphus antler keratinocyte growth factor kgf gene cloning, sequence analysis and transcription pattern research.docx

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马鹿cervuselaphus鹿茸角化细胞生长因子kgf:基因克隆、序列分析及转录模式研究-cervuselaphus antler keratinocyte growth factor kgf gene cloning, sequence analysis and transcription pattern research

东北林业大学硕士学位论文AbstractAsaspecificmitogenforepithelialcells,KeratinocyteGrowthFactor(KGForFGF-7),theseventhmemberofFibroblastGrowthFactors(FGFs)family,couldinduceproliferation,differentiationandmotility,andenhanceepithelialwoundrepair.ToclarifythepossiblerolesKGFmayplayindeerantlerdevelopmentandregeneration,forthefirsttimeinthisthesis,KGFgene(AccessionNo.AY923858)wasclonedfrom90一daydeerantlerofreddeer(Cervuselaphus)bytwostepRT-PCRmethod.BothKGFDNAandaminoacidssequencesofhuman,reddeeLmouse,sheep,dogandpig,wereusedforfollowingbioinformaticanalysisandsemi-quantitativeRT-PCRmethodwasfurtherusedtoexplorespacialtranscriptionaldynamicsofsuchgrowthfactorsasKGF,IGF—I,IGF一2,BMP一2,BMP一4.Resultsshowed:(1)KGFgeneofreddeercontains585bpCDSregioninlength,whichcodesforaputative194Aa●polypeptide.(2)Bioinformaticsanalysisshowed:thelinearfitcurveofKGFgenetransitionandtransversionamongthesixspecies:Y=一0.44925+0.37278x(∥=O.86115);theMEtreesthatwerereconstructedbasedonKGFgeneandpolypeptidesequence,inducedthatcomparedwithotherthreespecies,thehereditarydistanceofreddeer,sheepandpigshouldbeclose(bootstrap86%),thisresultwasconsistentwiththefunctionaldomainsforecastresultofKGFgenesecondarystructure;Foranytwospecies,neitherisolectricpointorchargeatPH7.0ofKGFwasequal,thereinto,isolectricpoint(9.173)andchargeatPH7.000.706)ofhumanwaslowest,andthepigwashighest,9.436and12.733separately;genesearchingresults:basedon2696motifsinthePrositedatabank,KGFmotifswereanalysed.KGFincluded46motifs,thereinto,FGFfamilysignatures,signalpeptiderelatedsignatures,andmetabolismrelatedsignatureswereveryimportant.Inaddition,transcriptregulationrelatedsignatureswerealsoinvolved,suchasLeucinezipperpaRem,ZincfingerRING-typesignature,ZincfingerC2H2typedomainsignature,ZincfmgerPHD—typesignature.ItinducedthatKGFmayparticipateingeneregulationprocess,however,todatetherehavenotbeenanyevidencestosupportthisviewpoint;thetypesofKGFgenemotifsvariedindifferentspecies.ComparationofhKGFanddKGFshowedthattherewasa5-nucleotidasesignature1at10

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