A Fish Eye Out of Water Ten Visual Opsins in the Four-Eyed Fish, Anableps anableps 英文参考文献.docVIP

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A Fish Eye Out of Water Ten Visual Opsins in the Four-Eyed Fish, Anableps anableps 英文参考文献.doc

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A Fish Eye Out of Water Ten Visual Opsins in the Four-Eyed Fish, Anableps anableps 英文参考文献

AFishEyeOutofWater:TenVisualOpsinsintheFour- EyedFish,Anablepsanableps GregoryL.Owens,DianaJ.Windsor,JustinMui,JohnS.Taylor* DepartmentofBiology,UniversityofVictoria,Victoria,BritishColumbia,Canada Abstract The ‘‘four-eyed’’ fish Anableps anableps has numerous morphological adaptations that enable above and below-water vision.Here,asthefirststepinoureffortstoidentifymolecularadaptationsforaerialandaquaticvisioninthisspecies,we describe the A. anableps visual opsin repertoire. We used PCR, cloning, and sequencing to survey cDNA using unique primersdesignedtoamplifyeightsequencesfromfivevisualopsingenesubfamilies,SWS1,SWS2,RH1,RH2,andLWS.We alsousedSouthernblottingtocountopsinlociingenomicDNAdigestedwithEcoR1andBamH1.Phylogeneticanalyses confirmedtheidentityofallopsinsequencesandallowedustomapgeneduplicationanddivergenceeventsontoatreeof teleostfish.Eachofthegene-specificprimersetsproducedanampliconfromcDNA,indicatingthatA.anablepspossessed andexpressedatleasteightopsingenes.AsecondPCR-basedsurveyofgenomicandcDNAuncoveredtwoadditionalLWS genes. Thus, A. anableps has at least ten visual opsins and all but one were expressed in the eyes of the single adult surveyed.Amongthesetenvisualopsins,twohavekeysitehaplotypesnotfoundinotherfish.OfparticularinterestistheA. anableps-specificopsinintheLWSsubfamily,S180c,withaSHYAAfivekeysitehaplotype.AlthoughA.anablepshasavisual opsingenerepertoiresimilartothatfoundinotherfishesinthesuborderCyprinodontoidei,theLWSopsinsubfamilyhas twolocinotfoundincloserelatives,includingonewithakeysitehaplotypenotfoundinanyotherfishspecies.A.anableps opsinsequencedatawillbeusedtodesigninsituprobesallowingustotestthehypothesisthatopsingeneexpression differsinthedistinctventralanddorsalretinasfoundinthisspecies. Citation:OwensGL,WindsorDJ,MuiJ,TaylorJS(2009)AFishEyeOutofWater:TenVisualOpsinsintheFour-EyedFish,Anablepsanableps.PLoSONE4(6): e5970.doi:10.1371/journal.pone.0005970 Editor:JohnWelch,UniversityofEdinburgh,UnitedKingdom ReceivedMay6,2009;Ac