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Oxford Genetics/pSF-Ad5 (OG268) Adenovirus Serotype 5 Plasmid/OG268/1 Ea
PlasmidInfo:
PlasmidInformation
ProductName:pSF-Ad5
ProductCode:OG268
Size(bp):33154bp
BacterialAntibioticSelection:
OriginandCompatibility:
BacterialCopyNumber:
Promoter:
PlasmidPurpose:
ThisisaSEROtype5adenovirusvirusvectorthatallowstheinsertionofgenesintotheE1Apositionwithinthevirus.ThevectorcontainsanE1andE3deletionandhasbeenmodifiedtoremoveanumberofkeyrestrictionsites.TheplasmidcontainsasmallmultiplecloningsiteconsistingofAsiSIClaIandPacI.Thesesiteswillbefoundinalloftheplasmidvectorsinourproductrangewhichallowsthedirectcloningofsegmentsfromourcatalogueplasmidsintotheadenovirusgenome(seecloningtabformoredetails).
PackagingCapacity:Thewildtypevirusgenomeis35938basepairsbutinthisvectorthegenomehasbeenshortenedbythedeletionstomakeit30501basepairs.Thisallowsfortheinsertionofupto5.5kbofexogenousDNA.Ithasalsopreviouslybeenshownthatserotype5adenovirusgenomescanaccomodateinsertionsupto105%totalgenomelengthcomparedtowildtype.ForthisreasonitshouldbepossIBLetoinsertanadditional1.7kbresultinginatotalinsertionsizeof7.2Kb.Wehavepreviouslypackagedvirusesupto105%ofthevirusgenomewithgoodstABIlitybutsomeinsertionsmaybeunstableatthislengthdependingonsequence.Thesenumbersarethereforeprovidedasguidelinesonly.
RecoveryandGrowth:VirusrecoveryrequirestwostepsfirstlytheplasmidmustbecleavedwiththeSwaIrestrictionenzymetolinearisethevirusgenome.Thenthelineargenomemustberecoveredandgrownin293cellsoranyderivativeof293cellssuchas293Asand293Tcells.TheselinescontainandexpresstheadenovirusE1Aproteinthattrans-complimentsthedeletionintheE1Aregioninthisplasmid.Atypicallyrecoverytransfectionwoulduseapproximately5ugofDNA.ItisnotnecessarytoremovetheplasmidbackboneafterlinearisingtheDNAastandardDNAcleanupwillbesufficientratherthangelextraction.Viruscoloniesshouldappearapproximately10-15daysaftertransfectioninto293cellshoweverclonescansometimesappearatearliertimepoints.Cellshouldbeculturedin2%fetalcalfserumtopreventovergrowthduringthistime.
SequenceandMap:
OtherInfo:
TranscriptionTermination:Notapplicable
IPStatus:
IntellectualPropertyStatusThisproductispartofourSnapFastProplasmidrange,formoreinformationontheIntellectualpropertystatusofthisplasmidpleaseclickhere
