Cloning and Partial Characterization of Tetrahymena thermophila Mitogen Activated Protein Kinase 3 (TtMPK3) Gene*

Journal of Applied Biological Sciences

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