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Oxford Expression /BacPAK6 Sec+/101104/5 reaction kit

Description:

BacPAK6Sec+linearizedbaculovirusDNAisthelatestbaculovirusexpressionsystemtocomeoutoftheOETlabs.BytakingtheoriginalBacPAK6systemandgeneticallymodifyingit,BacPAK6Sec+isabletoincreasetheyieldofrecombinantproteinandtheeaseofharvesting.

BacPAK6Sec+containsadeletionofthechitinasegene(chiA)fromtheoriginalBacPAK6viralbackbone.

Chitinasehasbeenshownbybothconfocalandelectronmicroscopytolocatetotheendoplasmicreticulum,whereitisdenselypackedinaparacrystallinearray,whichseverelyinhibitsthefunctionandefficacyofthesecretorypathway.BydeletingthechiA,secretionoftargetproteinsintothecytoplasmisincreased,greatlyenhancingtheyieldofharvestablerecombinantprotein.

TheconceptofdeletingthechiAgenecameafterthesuccessoftheflashBAC™platformthatutilizesthesamegeneticdeletionforincreasedrecombinantproteinyield.TofindoutmoreabouttheflashBAC™system,clickhere.

BacPAK6linearDNAistheoriginalconvenient,highlyefficientreagentforgeneratingrecombinantbaculovirusexpressionvectors. Itcomprisesamodified Autographacalifornica nucleopolyhedrovirus(AcMNPV)genomewith lacZ insertedinplaceofthenativepolyhedringenecodingregionforconvenientblue/whiteselection.

DigestionofBacPAK6DNAatthreesiteswithBsu36Iremoves lacZ andpartofORF1629,whichisanessentialAcMNPVgene. ThelinearvirusDNAisunabletoinitiateareplicationcycleininsectcells.
Co-transfectionofinsectcellswithlinearBacPAK6DNAandacompatIBLetransfervectorcontainingthecompleteORF1629andaforeigngeneofinterestresultsinrecombinationandrestorationofthecircular,infectiousvirusgenome. Over95%ofthevirusderivedfromtheco-transfectioncomprisesrecombinantvirus. Usually,asingleplaqueassaytitrationincubatedwithX-galandcounterstainedwithneutralredissufficienttodifferentiateparentalbluefromrecombinantwhite(colourless)plaques. ThesecanbereADIlyisolatedandamplifiedtoworkingstocksofpurifiedrecombinantvirusinafewdays.

TheBacPAK6Sec+reagentfromOETLtdcomprisestransfection-ready,linearized(Bsu36I-digested)virusDNA. Itisavailablein5,24and96reactionpacks. Forbulkpurchases,pleasecontactusoninfo@oetltd.com.

WeofferapromotionaldealonallBacPAK6productstoincludeafree3reactionflashBAC™kittohelpyoudecideifmakingtheswitchistherightchoiceforyourlab.

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