Description:
pTWIN1isanE.coliexpressionvectorwhichcanbeusedwiththeIMPACT™Kit(NEB#E6901).pTWINvectorsaredesignedforproteinpurificationorfortheisolationofproteinswithanN-terminalcysteineand/oraC-terminalthioester(1).Apolylinkerinthevectorisdesignedforthein-framefusionofatargetgenebetweenthemodifiedSspDnaB(2)andMxeGyrAinteins(3,4).ThepresenceofthechitinbindingdomainfromBacilluscirculans(5,6)facilitatespurification.Thedouble-strandedvectoris7,375basepairsinlength.
DNASUisacentralrepositoryforplasmidclonesandcollectionsthatmayalsobehelpful.
Notes:
CellLysisBuffer:50mMTris-HCl(pH8.5)containing500mMNaCl.SspDnaBInteinCleavageBuffer:50mMTris-HCl(pH6.0)containing500mMNaCl.MxeGyrAInteinCleavageBuffer:50mMTris-HCl(pH8.5)containing500mMNaCland50mM2-mercaptoethanesulfonicacid.