Description

pOET2.1N/C 6xHis (cat# 2001031) (10ug) is a baculovirus transfer vector designed for high level expression of foreign genes under the powerful AcMNPV polyhedrin (polh) promoter.  The vector encodes an N-terminal 6xHis-Tag^® fusion sequence that may be utilised if the insert includes a stop codon.  This greatly eases the purification of the recombinant protein since the 6xHis-containing fusion proteins bind with high affinity to Ni-NTA Agarose.  If required, the 6xHis-Tag^® can be removed by incubating the fusion protein in the presence of the proteinase cleavage enzyme Thrombin. There is also a 6xHis-Tag^®for C-terminal fusions where the insert’s own start codon can be used to replace the start codon supplied in pOET2.1N/C. pOET2.1N/C is smaller than other available transfer vectors (4653 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 The polh sequences have been replaced by a multiple cloning site (MCS) containing unique restriction sites for insertion of the foreign gene in the correct orientation. The AcMNPV sequences flanking the gene in the transfer vector’s MCS allow recombination with the viral DNA to insert the expression cassette into the polh locus. pOET2.1N/C is compatible with any baculovirus system that utilises homologous recombination in insect cells.

Variations

• pOET1.1N 6xHis has the same MCS as pOET2.1N/C 6xHis except in the reverse orientation.
• pOET2.1C 6xHis has a C-terminal rather than N-terminal 6xHis-Tag^®.