Journal of Applied Biological Sciences | |
Cloning and Partial Characterization of Tetrahymena thermophila Mitogen Activated Protein Kinase 3 (TtMPK3) Gene* | |
Muhittin ARSLANYOLU1  | |
关键词: Temperature downshift; MAPK; cDNA; mitogen; ciliate; unicellular; | |
DOI : | |
学科分类:生物科学(综合) | |
来源: Nobel Bilim ve Arasirma Merkezi/Nobel Science and Research Cente | |
【 摘 要 】
Inthisstudy,afull-lengthcDNAfragmentencodingputativemitogenactivatedproteinkinase(MAPK)fromTetrahymenathermophilawasclonedbyusingRT-PCRwithdegenerateoligonucleotideprimers.Baseduponthepresenceof11subdomainsandTEYdualphosphorylationmotifinproteinkinasesubdomainVIII,itisnamedasTetrahymenathermophilamitogenactivatedproteinkinase3(TtMPK3).TtMPK3cDNAfragmenthas1176bplongsingleopenreadingframe(ORF)encodingapolypeptideof391aminoacidswithpredictedsizeof45.5kDa.TtMPK3ishighlyhomologoustoSLT2ofbuddingyeastandErk1ofGiardiaintestinalis.SouthernblotanalysisindicatedthatTtMPK3isasinglecopygeneinTetrahymenamacronucleargenome.Westernblotanalyseswithanti-carboxylTtMPK3ppeptideserumshowedthatTtMPK3polyclonalantibodyspecificallyrecognizesonlyabout44kDaexpectedsizeprotein.Immunoprecipitatesofanti-carboxylpolypeptideofTtMPK3pantibodytestedininvitrokinaseassayshowedkinaseactivitytwotofourtimesmorethanthecontrolpreimmuneserum.Untiltoday,fourpossibleTtMPKgenesincludingTtMPK3wereidentifiedandtheTetrahymenaGenomeProjectidentifiedadditional11membersfromp38ErkandErk7subgroupofMAPKfamilyinTetrahymenagenome.
【 授权许可】
Unknown
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