Escherichia coli K12 a cooperatively developed annotation snapshotWord格式文档下载.docx
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2DepartmentofGenetics,CandidaGenomeDatabaseStanfordUniversitySchoolofMedicine,Stanford,CA94305-5120,USA
3DepartmentofMolecular,CellularandDevelopmentalBiology,YaleUniversity,NewHaven,CT06520-8103,USA
4GenomeCenterofWisconsin,425HenryMall,UniversityofWisconsin,Madison,WI53706,USA
5DivisionofImmunityandInfection,UniversityofBirmingham,Edgbaston,BirminghamB152TT,UK
6NationalInstituteforBasicBiology,Nishigonaka38,Myodaiji,Okazaki444-8585Aichi,Japan
7SRIInternational,333RavenswoodAvenue,MenloPark,CA94025-3493,USA
8GraduateSchoolofBiologicalSciences,NaraInstituteofScienceandTechnology,Ikoma,Nara630-0101,Japan
9TheInstituteofAdvancedBiosciences,KeioUniversity,Tsuruoka,Yamagata997-0017,Japan
10DepartmentofBiochemistryandMolecularBiology,TheUniversityofMiamiMillerSchoolofMedicine,Miami,FL33140,USA
11DepartmentofBiology,UniversityofYork,POBox373,YorkYO105YW,UK
12TheWellcomeTrustSangerInstitute,GenomeCampus,Hinxton,CambridgeCB101SA,UK
13DepartmentofComputingScienceandBiologicalSciences,2-21AthabascaHallUniversityofAlberta,Edmonton,Alberta,CanadaT6G2E8
14DepartmentofBiologicalSciences,PurdueUniversity,915W.StateStreet,WestLafayette,IN47907-2054,USA
*Towhomcorrespondenceshouldbeaddressed.Tel:
+15082697388;
Fax:
+15084574727;
Email:
mriley@mbl.edu
CorrespondencemayalsobeaddressedtoBarryL.Wanner.Tel:
+17654948034;
+17654940876;
blwanner@purdue.edu
ReceivedNovember5,2005;
RevisedDecember5,2005;
AcceptedDecember5,2005.
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Abstract
INTRODUCTION
RESULTSANDDISCUSSION
RESULTS
SUPPLEMENTARYDATA
SupplementaryMaterial
REFERENCES
Abstract
Thegoalofthisgroupprojecthasbeentocoordinateandbringup-to-dateinformationonallgenesofEscherichiacoliK-12.Annotationofthegenomeofanorganismentailsidentificationofgenes,theboundariesofgenesintermsofprecisestartandendsites,anddescriptionofthegeneproducts.Knownandpredictedfunctionswereassignedtoeachgeneproductonthebasisofexperimentalevidenceorsequenceanalysis.Sincebothkindsofevidenceareconstantlyexpanding,noannotationiscompleteatanymomentintime.ThisisasnapshotanalysisbasedonthemostrecentgenomesequencesoftwoE.coliK-12bacteria.Anaccurateandup-to-datedescriptionofE.coliK-12genesisofparticularimportancetothescientificcommunitybecauseexperimentallydeterminedpropertiesofitsgeneproductsprovidefundamentalinformationforannotationofinnumerablegenesofotherorganisms.AvailabilityofthecompletegenomesequenceoftwoK-12strainsallowscomparisonoftheirgenotypesandmutantstatusofalleles.
INTRODUCTION
EscherichiacolistrainK-12isarguablythesingleorganismaboutwhichthemostisknown.Originallyisolatedin1922,itwascatapultedtoprominencebythediscoveryofstrainK-12'
sabilitytocarryoutgeneticrecombinationbyconjugation
(1)and,soonafter,bygeneralizedtransduction
(2).ThestrainK-12hasbeenwidelydistributedtolaboratoriesacrosstheworld.Overtheensuingyearsitbecametheprimarymodelorganismforbasicbiology,moleculargeneticsandphysiologyofbacteria,andwasthefoundingworkhorseofthebiotechnologyindustry.
AnnotationofE.colihasnotonlyservedtheE.colicommunity,buthasformedabasisforextrapolationofgenefunctionstovirtuallyeveryotherprokaryotic,aswellaseukaryotic,genomethroughanalogybasedonproteinsequencesimilarities.Assuch,theaccuracyandcompletenessoftheE.coliinformationisofgreatimportancetothecommunityofbiologistsworkinginalldisciplinesandwithallorganisms.WereportheretheworkofagroupofscientistsdedicatedtofullreviewandupdateoftheannotationofE.coliK-12.
TheentiregenomesequenceofK-12strainMG1655wasfirstcompletedandannotatedbyagroupassembledbyF.R.Blattner(3).ThegenomeofasecondK-12strain,W3110,wascompletedrecentlyunderthedirectionofTakashiHoriuchiattheNationalInstituteforBasicBiologyinJapan(4).AtthesametimethesequenceofthegenomeofMG1655wascorrectedandupdated.MG1655waschosenforitscloserelationshipwiththeoriginalE.colistrainK-12(calledEMG2),whereasW3110waschosenbecauseithasbeenwidelyusedasa‘wild-type’strainbymanyinvestigationsworldwidefromthe1950s.BothhadbeencuredoftheλprophageandlacktheF+fertilityfactorofancestralE.coliK-12EMG2.MG1655andW3110are1-and2-stepdescendentsofE.coliK-12W1485(F+,λ−),respectively,whichisinturnadirectdescendentofEMG2(4,5).
Bycomparingandre-sequencingregionsofdiscrepanciesbetweenMG1655andW3110,highlyaccurategenomeshavenowbeencreatedforbothstrains(4).CorrectionstotheoriginalMG1655genome(3)areat243sites(totaling358nt),acorrectionrate8yearslaterof~7in105.WorkdonebytheparticipantsofanE.coliannotationworkshopheldinNovember2003reconciledsequencedifferencesthatledtodepositofacorrectedMG1655genomesequenceentry(GenBank™U00096.2,releasedinJune2004).SubsequentworkdoneinaMarch2005workshopintroducedadditionalchanges.Theparticipantsoftheseworkshopshaveco-authoredthismanuscript.
AlthoughbothMG1655andW3110areisolatesoftheE.coliK-12strain,theirgenomesarenotidentical.ThedifferentlengthsoftheMG1655(4639675nt)andW3110(4646332nt)genomesreflectalargernumberofinsertionsequence(IS)elementsandabsenceofadefectivephageintheW3110genome.Otherdifferencesarefoundintheoccurrenceofmutations,reflectingchangesthatpresumablyoccurredduringmaintenanceoftheculturesinseparatelaboratories.
Genomeannotation,ofnecessity,isanongoingprocess.Intheinterimfrom1997,manyscientists,notorganizedasagroup,butunitedintellectuallybytheirinterestindevelopingaunifiedvisionoftheorganism,havecontinuedtoupgrade,updateandcollatenewinformationaboutE.coliasithasemerged.Thishasresultedinanumberofpublicdatabaseswithinformationongenes,genomicsandproteinsofE.coliK-12,noneidentical,eachwithadifferentemphasis.Othermoregeneraldatabasescontaininformationrelevanttomanyorganisms,helpfulininterpretationofgenesequences.
ThegoalofthecurrentprojectwastoconsolidatetheworkofscientistswhohavebeenworkingindependentlybydevelopingourbestconsensusonthestatusandpropertiesofeachofthegenesofE.coliK-12atthepresentmoment.Thegoalwasdecidedlynottocreateanewdatabase,butinstead,topresenttothepublicacomprehensive,updatedannotationofE.coliK-12whichwouldbepresentedbothinspreadsheetandsimpleflat-fileformats.Thelattercaneasilybeparsedbycomputersandreadersalikeandthereforecanbeincorporatedintoextantdatabasesbytheirproviders.TheseareavailableasSupplementaryTable1.xls,SupplementaryTable1.txtand,toaidininterpretingthedata,SupplementaryTable1ExplanatoryNotes.LessextensiveinformationfromthenewMG1655andW3110annotationshavebeenincludedinnewGenBank™andDNADataBankofJapan(DDBJ)entries,accessionnumberU00096.3andDDBJAP009048,respectively.
Werefertothisoutcomeasa‘snapshot’toemphasizethatinformationaboutE.coligenesandtheirproductsareamovingtarget,andovertakenrapidlywithmorerecentinformation.Theauthorshavemadenoplanstodevelopthissnapshotfurther.HighlydesirablewouldbetheestablishmentofanaccessiblecommunityresourceofdataonE.coliK-12withcommunityparticipation,ongoingmaintenanceandcontinuousupdatingofallinformation.AtthismomentinterestedmembersoftheE.colicommunityareapplyingtoNIHforsupporttoestablisha‘K-12informationresource’.
RESULTSANDDISCUSSION
Theworkshops
Theneedtoconsolidatetheeffortsofscientistswhohadbeenworkingindependentlywasasubjectofdiscussionataninformal‘E.coliconsortium’meetingorganizedbyBarryWanneran