Oxford Expression Technologies

pOET2N/C_6xHis is a baculovirus transfer vector designed for high level expression of foreign genes under the powerful AcMNPV polyhedron (polh) promoter.  The vector encodes an N-terminal 6×His-Tag^® fusion sequence that may be utilized if the insert includes a stop codon.  This greatly eases the purification of the recombinant protein since the 6×His-containing fusion proteins bind with high affinity to Ni-NTA Agarose.  If required, the 6×His-Tag^® can be removed by incubating the fusion protein in the presence of the proteinase cleavage enzyme Thrombin. There is also a 6×His-Tag^®for C-terminal fusions where the inserts own start codon can be used to replace the start codon supplied in pOET2N/C_6xHis. pOET2N/C_6xHis is smaller than other available transfer vectors (4626 bp) which greatly facilitate the cloning steps. It has a Col E1 origin of replication and an ampicillin resistance gene for selection in E. coli. The polh sequences have been replaced by a multiple cloning site (MCS) containing unique restriction sites for insertion of the foreign gene in the correct orientation, as shown on the circular map.  The coding strand of the MCS as transcribed from the polh promoter is shown below the circular map.  The PacI site at the end of the MCS provides translational stop codons in all three reading frames for expression of truncated proteins. The AcMNPV sequences flanking the gene in the transfer vectors MCS allow recombination with the viral DNA to insert the expression cassette into the polh locus. pOET2N/C_6xHis is compatible with any baculovirus system that utilizes homologous recombination in insect cells.

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