Designation:E1493–06StandardGuideforIdentificationofBacteriophageM13orItsDNA1ThisstandardisissuedunderthefixeddesignationE1493;thenumberimmediatelyfollowingthedesignationindicatestheyearoforiginaladoptionor,inthecaseofrevision,theyearoflastrevision.Anumberinparenthesesindicatestheyearoflastreapproval.Asuperscriptepsilon(´)indicatesaneditorialchangesincethelastrevisionorreapproval.INTRODUCTIONThisguidecoverstheidentificationofbacteriophageM13oritsDNAandwasdevelopedbyTaskGroupE48.02.03onviruses.TheobjectiveistodescribelaboratorycharacterizationproceduresthatwouldbesufficienttoverifythatabiologicalpreparationbelievedtocontainM13(orM13DNA)foruseinanystepofabiotechnologyprocessactuallydoescontainthisbacteriophage(oritsDNA).Thisguideassumesabasicknowledgeofmicrobiologyandmolecularbiology.1.Scope1.1ThisguidecoverstheidentificationofbacteriophageM13usedinbiotechnology.1.2TherearemanyvariantsofM13thathavebeendevel-opedspecificallyforcloningtechnology.ThesevariantshaveforeignDNAinsertedintotheM13genome,causingtheM13todifferinsizeandgenotype.1.3IftheM13istobeusedtoconstructarecombinantmolecule,thenthecriteriadescribedinSection6shouldbeusedtocharacterizethenewlymadeDNA.2.Terminology2.1Definitions:2.1.1alphacomplementation—theabilityofashortamino-terminalfragment(alphafragment)ofb-galactosidasetoformafunctionalcomplexwiththecarboxylterminalfragment(omegafragment).2.1.2F1—anFfactorthatcontainsaportionoftheE.coligenome.2.1.3F+factor—anepisomeofE.coli.EncodedonitarethefunctionsnecessarytoproduceanFpilus.2.1.4Fpilus—aprotrusiononE.colithatisnecessaryformating.TheFpilusalsocontainsthereceptorforphageM13.2.1.5Hfr—astrainofE.coliinwhichtheF+factorisinsertedintothechromosome.2.1.6multiplecloningsite—DNAthatcontainsseveralcontiguousrestrictionenzymerecognitionsites;alsocalledapolylinker.2.1.7phagedisplay—theuseofbacteriophageM13oravarianttoinsertapeptidesequencefusedtoacoatproteinofM13.3.SignificanceandUse3.1Thisguideisintendedforuseinabiotechnologylaboratorywhentheneedarises...
