3477

3478

9000001143733

Hirumi, Hiroyuki

Majiwa, P.

Urakawa, T.

English

OAK Obihiro University Archives of Knowledge

．／．／’化けりこ〔ノ〃／．J（ぐ．I．、お，㍊4ヱ2βりリリ叫  C叩Y＝．？加¢J9相，川ピ鮎†印mH∴～′けどr♪汀什りJ√）＝√′一7〃1わ／打′両′■／〝＝7川／7り／r代l、  Compamtive ana］yses ofrihosomalRNA genes of●AfticaLlaIldrelated  trypanosomes，imcltIding rりpα乃〃∫♂mαぞyα〝∫f  T．URAKAWAl，P．MAJIWA2andll．HIRUMILl  l∫√′（・k丹♪〝r汀血相f′～′′〔川〃仙〃可■．4grん・扉〃′rど，抽rど∫呵‘川‘肌血r如いイン1什り′∫～〟川／G率帖」  1押〃＝血んハ〟b血，仙汀√粛J〃5イ棚4，ノ叩岬コ加ビr′刷／（川√J／J一雨J・J（′（・川ビJ（ノ√′両～ノ什～J加J‘ノ，Jl（ノ．β…・  ．す07瑚〃肌r赫，放′岬，377ば肋ピ‘′rぐん〔加／〝ノわr什0／（′こ〃√川肱血・∫血rJ川′′川′∫叫どV，（ノ肋血U′′血r叫  （ゾAgrmJ／川r亡用／Jr／l々／ぐrf／汀汀Y．W（ノ／／ノ川J（一J†J／J（／（汀血イル′／〃〝J，〟／ノ抽Jんわ†Jβ（用5∫5‥〟叩JJ  Re（■C‖ハHlユ（）′＼∪旨u、1川リ8  Molecularphylogeny providesanalIemativeviewon evolutionaryrelalionships among  relatedorganisms．Thecomparisonofthesequencesoftheribosoma［RNAgenes（rDNAs）  isapowerfultoolforphyLogene（icanalysis・TherDNAsareoneofthemulticopygenesand  aretandemlyrepeatedinthegenome（Fig・1）・  コ11  l・烏射  】00124  16p  5る0＿ 5る三  r．α叫卯血徽陀S馴伯l郎血・typモ  ．▲＿＿ ■¶ l．．．n  r√朋g（7血刀∫g WA∬・けpモ   rf〃〃g（〉Jg〃gg鮎l漬・けpモ  r“明画加朋Ts且Y｛l・lype  r．∫f椚血ゼ  r．yル丘∫  r．1hnル¶  J■、rlu‖tl  r♪川r■l恒¶．・′l  三Ⅲ   豊根  21壬  ユl】   七＝＝ 】69  l朗  1．呈‡当 ＿ヱウ9＋  】b9 t tL竺p■， 【TS：Elt甘n●lTr●叩CrlbdSp…r  】TS：lnlt▼n81Tr■n虻rlMS叩¢℡r  yRN▲：Oml州－1血01n■lRll▲  28S  Figure．1．Thegenemapsord11brenL LT？；PinOSOmCSinc）udlng7万，PLu，OL甘けn〟eVr〝1SiL Allsquence血軋  ？XCePtlbrT・bTu（－eibTu（leE，WCrCObtalnedinthisstudy・ThehbosomaLRNAgenes（rDNAs）arcTCfXntCJ  lnlancbn皿d∬etranSCnbedasa】ongprecursorRNAmoleiruLcstartLngrrOrnthctTuSCnPtlOnlnLtlirtion  Si【ein（hepromoLersequence・Then thc LTⅦSulPtis cleilVedimmedlatdy jnto18S－5・8S∴撼SLSL  28S－LS2，andsmal1rlbosoma）RNAsbythespeCjricRNases・   TheunlqueCharacteristicofrDNAsisexplainedbytheconcertedevolution・According  tolhe concerted evolution，geneS are homogenized during evolution of species by  homologousrecombination，unequalcrossoyer・Sisterchromatidexchange，geneCOnVerSion  or other mechanisms．In order toinvestlgate queStlOnS SUrrOunding the phylogenetlC  reIationshipsamong African trypanosomes，We haveanalyzedrDNAsof31strainsofll  differenttrypanosomes：T［ypan（）Snmael，an・9f（8s（rains）†r・b・bruceL（2），T・h・gambi  （7），Tlb．rhodesiense（2），TIcongoLense［savannal1－tyPe（5），WestAfrican－riverinepforest  type（1），Ki1ifi－type（1），Tsavo－tyPe（1）」，I二vivax（1），Trsimlae（1）andT・thei／eri（2）・The  ribosomalRNAgenes，including18Srt）NA，5■JllS，5・8SrDNA，3rlTS，and28S－LSl，Were  ∴‘   RIBOSOMALRNAGENt：SOFAFR（CANTRYPANOSOMES  obtained either as genomic DNA clonesinlambda phage or as PCR products from  trypanosomcgenomicDNA，andwereeithersequenceddirectlyorsub－CIonedintopIasmid  vectorspr10rtOthesequcnclng・ThephylogenetictreeswereobtainedbydifferentanalysIS  programs（Fig・2）・  Figure2．The phylogeEleticLreeOr18S－DNA・Thc（recwasobtaJnedby asmalt compllterPJDgmm，  CLUSTAL W（Ver，1，74，ThompsonJD，HiBgir）S，DG，肌d Gibson TJ：Nucleic Aclds Res，22，  4673欄0・1994）・F（一urty甲OrT・COTLgOle旧eWeTelo侃tCdasdLfferenlb一ⅦChes二Incontrasttothis，  thes叩1enCe d胞陀nO防ⅥⅦ℃n（－tObs肌山地w∝Ilr－・どy‘mf諷dr・わ川α～占mαJincludng r・わ川αi  脚鵬血Ⅳ即dr・わr7灯girん血・1■鹿丑貯一  Weflrst analyzed the rDNAsof four differenttypeS Of TIcongoLense．There was  SIBnificantheterogeneltyamOngthe sequences，makingeach ofthe trypanosomescLearly  distinguishableonthisbasis．ThesequencedifferenceswereobservedinthespacerreglOnS，  the18S and the28S－LSlrDNA scgmetlts．ATnOng8isolatesofsavannah－tyPefromthe  differentpartsofAfrica，anjmsignificantnumberofnucleotides11bstitutionsl嘩reObserved  in the18SrDNA region・Of the different typesof7二congL？LeTISe（Accession Number：  U22315），theWestAfrican－riverine－forcsttype（U22319）wasfound to beclosesttothe  SaYannah－tyPe．With97％matches，fbllowedbytheKilifi－type（U22317）with93％matches－  Figure3．An applicalion fordLagnOSIS tO虚5tinguish s匹CleS OrtypeS OrtryPanOSOme＄uSlng rDNA  S叩enCe血はmePCRampliriG血onorthe陀糾OnOr5’ETS（extem血lⅧ即納d叩舶両町dSouth訂n  blotanalysiswereshown．SixtemDNAsampleswe托amPLifiedbyPCRusingap剋rOrPnmerSWhidl  amplifies550bpin51ETS orT・mngOLenseSayannah－ty匹and44ObporTIcongoねnseKllifi－type・A＝  2％agamsegelcleetrophorcsisofthePCRpl血ICtS・8＝SouthemblotpIDbedwith700bpETSDNÅh）m  theSavanndl－1ype・ThesamPlenumbcrl，2，and8weretheSavannah－type・Thesamplenumber16were  mixturcoftheSavannahrtypeandtheKl］ifi－type．TheothersweretheKilifi一（ype－  ∴、j   RIBOSOMALRNAGENESOFAFRICANTRYPANOSOMES  Curiously，theTsavo－type（U22318）T，C（フng（，［ensewasphylogeneticallyc］osertoT・1Timiae  （U22320）thanit was to the savannah－type．Forinstance，We COuId differentiate the  Savannah－tyPe and（he KilifLLype with ease usLng the differences of rL）NA sequences  （Fig．3）．Ourdatastronglysuggeststhatthefourtrypanosomesnowgroupedtogetheras71  COngO／en5eCOuldt光ClassifiedintoindividualspecleS・  Thissituation contrastswithwhatweobservedin theanalysIS Of7二Jwuceiand related  trypanosomes，The subgenus7bpan（）こ0（）n COmprlSeSthree speCies＝T・brucei，7t et，anSi  （U75507，D89527）andTequiT，erdLi肌・WthinT・bruceithreearesubspecies：Tlb・bruFei  （M12676），Tlb．gambiense andT・b・rh〃desiense・In additionto hostrange，the m叫Or  phenotypic difference among trypanosomesin（his subgenusis the absence of kDNA  maxicircle、Whichiscorrelatedwiththecapacltyforcyclica］developmentinthetsetsefly・  Wefoundafewnuc）eo（idcbascsubstitutionsirtbo（hthe18SrDNAandthespacerregIOrtS  whentherDNAscquencesofthesetrypanosomeswerecompared；Wehadnodatafor71  eqELOerdELm．1tisthoughtthat7二evan5ievoIvedfromT・bruL・el，and（henspreadfromAfrica  tootherpartsoftheworldin（herecentpast・AlthoughtTletWOtryPanOSOrneShavedifferent  lifestyles，We found no evidence from the anaJyses of rDNA genes to support their  classificationin10different speCies evenin the spacer sequences・For simpIiclty from  evolutionarypoLntOfviewtheymayt光regardedassubspeciesorvarlantSd■T・brucei・  Inaddition，itbecamecIearthatTvivax（U22316）waslocatedoutofboththeSalivarian  andStercoてariangroupsandT・theiLeri（ABOO7814）wasclosetoT・Cruこi（M31432）inthe  phylogenetlClreeof18SrDNA（Fig・2）・   REFERENCIミS  Chmpt｝ell，D・A・，Kubo，K・，Clark，C・G・＆BoothroydJ・C・1987・Preciseidentificationof   cleavagesitesinvoIvedintheunusualprocesslngOftrypanosomeribosoma］RNA・   J．AわJ．郎oJ．ノ5婚ご／Jj－ノ2‘′．  Lukes，，．，Jirku．M．，Dolezel，D．，Kml－ova，Ⅰ．，Ho11ar．L．＆Mas）ov，D．A．J997．AnalysISOf   ritx）SOmalRNAgenessuBgeStSthattrypanosomesaremonophyletic・J・眈，l・Etd・   ∫2ト∫27．  Maslov、D・A・＆Simpson，L・1995・Evolutionofparasitisminkinetoplastidprotozoa・   Pαrα∫加J．m（わyJノJjO－j2．  Maslov，D．A．，Lukes，J．，Jirku、M．＆Simpson，L・1996・PhyLogenyoftrypanosomesas  inferredfromthesmal1andlargesubunitrRNAs：implicationsfortlleeVOlutionof   parasi（isminthetrypanosomatidprotozoa・朗bl・Biochem・Partzsifo／・75：197－205・  ThompsonJ・D・．Higgins，D・G・＆GibsonT・J・1994・CLUSTALW：improv．ngthe   sensitivityofprogressivemultiplesequencealignmentthroughsequenceweighting，   position－SpeCificgappenaltiesandweightmatrixchoice・NucleicAcidsRes，22：   467j一イ∂β0．  White，T．C．，Rudenko，G．＆Borst，P．］986．TreesmallRNAswlthjnthe10kbtrypanosome   rRNAtranscriptlOnunitareanaJogoustodomainVIlofothereukaryotic28SrRNAs・  〃〟CJgfcA（－i〟∫月ビ∫．ノ4．・タイ7ノー9〃βp．  二■ノト   

Comparative analyses of ribosomal RNA genes of African and related trypanosomes, including Trypanosoma evansi

https://obihiro.repo.nii.ac.jp/record/1576/files/Prot.Vol.8-4-3.pdf

Comparative analyses of ribosomal RNA genes of African and related trypanosomes, including Trypanosoma evansi

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