pOET2N/C_6xHis昆虫杆状病毒表达载体-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心

pOET2N/C_6xHis昆虫杆状病毒表达载体-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心

Plasmid Description:

pOET2N is a baculovirus transfer vector designed for high level expression of foreign genes under the powerfulAcMNPV polyhedron (polh) promoter. The vector encodes an N-terminal 6×His-Tag®fusion sequence that maybe utilized if the insert includes a stop codon. This greatly eases the purification of the recombinant proteinsince the 6×His-containing fusion proteins bind with high affinity to Ni-NTA Agarose. If required, the 6×HisTag®can be removed by incubating the fusion protein in the presence of the proteinase cleavage enzymeThrombin. There is also a 6×His-Tag®for C-terminal fusions where the inserts own start codon can be used toreplace the start codon supplied in pOET2N. pOET2N 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 genefor selection in E. coli. The polh sequences have been replaced by a multiple cloning site (MCS) containingunique restriction sites for insertion of the foreign gene in the correct orientation, as shown on the circularmap. 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 forexpression of truncated proteins. The AcMNPV sequences flanking the gene in the transfer vectors MCS allowrecombination with the viral DNA to insert the expression cassette into the polh locus. pOET2N is compatiblewith any baculovirus system that utilizes homologous recombination in insect cells.

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