pex-seriesofprecisionshuttledestinationvectors.docVIP

pEX-series of PrecisionShuttle Destination Vectors For Protein Expression in E.coli pEX-N-His-GST pEX-N-GST pEX-N-His pEX-C-His pEX-1 Introduction The pEX-series of vectors are destination vectors for protein expression in E. Coli. The T7 promoter in the vector allows the encoded protein to be expressed in BL21(DE3) cells upon IPTG induction. The pEX-series of vectors also provide a TEV (the tobacco etch virus) protease cleavage site to remove any of the tags after protein purification. The pEX N-terminal tagged vectors share the same multiple cloning sites as other destination vectors to facilitate easy transfer of the ORF region from any of OriGene’s standard TrueORF clones in the pCMV6-Entry vector. pEX-1 and pEX-C-His vectors have a BseR I site instead of Sgf I due to the restriction of E. Coli RBS (ribosome-binding site) for higher protein expression. BseR I and Sgf I produce compatible sticky ends, so pEX-1 and pEX-C-His vectors are still compatible with transferring the ORF region from any of OriGene’s standard TrueORF clones in the pCMV6-Entry vector or other destination vectors. The PrecisionShuttle, a one-step cut/paste procedure, is fast, reliable, and cost effective. The different antibiotic resistance marker in the destination vectors (Ampicillin instead of Kanamycin in the Entry Vector) is designed for easy screening of the successfully shuttled plasmids. The diagram below shows how to do subcloning from TrueORF Entry into other destination vectors. Note: to clone an ORF into pEX-1 and pEX-C-His vectors, BseR I/Mlu I will be used instead of the typical Sgf I /Mlu I. The pEX vector series have several tagging options, tagged and non-tagged or on either the N- or C-terminus. Depending on the desired purification strategy or desired fusion partner, one can choose the N-His-GST-, N-GST, N-His-, C-His-tagged or non-tagged bacterial expression vector. For details, please see /cdna/TrueORF/DestinationVector.aspx , then click on the “Bacteria