Oxford Expression Technologies - pOET Transfer Vectors
E-mail Print
Information on the Oxford Expression Technologies pOET™ Tansfer Vectors
Go to Oxford Expression Technologies website
Oxford Expression Technologies has release a new family of Transfer Vectors that are designed to form the perfect complement to their existing flashBAC™ range of vectors. The gene of interest undergoes homologous recombination into the virus genome (eg  flashBAC™) from the transfer Vector. Hence cloning into the transfer plasmid forms the essential first step in a baculovirus expression experiment. The new pOET™ Transfer Vectors offer a number of features that make them an ideal starting point for expression work.
pOET Transfer Vectors
Go to Information on Compatibility of pOET with other Expression Systems Compatability of pOET transfer Vectors with other Baculovirus Expression Systems
Go to Technical Overview of pOET Transfer Vectors Technical Overview of pOET Transfer Vectors
Go to Information on pOET1 and pOET2 pOET1 and pOET2
Go to Information on pOET3 and pOET4 pOET3 and pOET4
Go to Information on pOET5 pOET5
Go to Information on pOET1C and pOET2C pOET1C_6xHis and pOET2C_6xHis
Go to Information on pOET1N and pOET2N pOET1N_6xHis and pOET2N/C_6xHis
Specific Product/Application Areas:
Go to flashBAC Product Information flashBAC Baculovirus Protein Expression
Go to baculoQUANT Product Information baculoQUANT Virus Titration
Go to titrePLUS Product Information titrePLUS Virus Titration and Protein Expression
Go to baculoFECTIN Transfection Reagent Information baculoFECTIN Transfection Reagent
Go to NEW Products NEW Products
Go to NEW superSf9 Cells NEW superSf9 Cells
Go to NEW Transfer Vectors Four NEW Transfer Vectors for C-Terminal and N-Terminal 6xHis-Tag® Fusion Sequences
Go to baculoFECTIN Transfection Reagent NEW baculoFECTIN Transfection Reagent
Go to NEW baculoGROW Serum-free Insect Medium NEW baculoGROW Serum-free Insect Medium
Go to NEW baculoCOMPLETE - ALL-IN-ONE Baculovirus Expression Kit NEW baculoCOMPLETE - ALL-IN-ONE Baculovirus Expression Kit
Go to NEW flashBAC™ Selection Box NEW flashBAC™ Selection Box
Go to NEW 96 Reaction flashBAC™ Kits NEW 96 Reaction flashBAC™ Kits
Literature
Go to Technical Literature Download Oxford Expression Technologies Technical Literature
Go back to Oxford Expression Technologies Products and Applications Oxford Expression Technologies Products and Applications
Compatability of pOET Transfer Vectors with other Baculovirus Expression Systems
pOET™ Transfer Vectors
Go to Oxford Expression Technologies website
pOET range of Transfer Vectors (Plasmids) are compatible with any baculovirus system that utilises homologous recombination in insect cells including the flashBAC products from Oxford Expression Technologies (OET). Below is a list of those baculovirus expression systems which are compatable with the pOET Transfer Vectors:
 
  • flashBAC (OET)
  • flashBACGOLD (OET)
  • flashBACULTRA (OET)
  • BacMagic (Novagen)
  • BacMagic2 (Novagen)
  • BacVector 1000 (Novagen)
  • BacVector 2000 (Novagen)
  • BacVector 3000 (Novagen)
  • BacPAK6 (Clontech)
  • BaculoGold (Clontech)
  • BaculoBright (Clontech)
  • ProGreen (AbVector)
  • ProFold (AbVector)
  • ProEasy (AbVector)
  • Sapphire (Orbigen)
Go to Top
Technical Overview of pOET Transfer Vectors
pOET range of baculovirus transfer vectors (plasmids) are designed for high level expression of foreign genes under either the powerful AcMNPV polyhedrin (polh) promoter or under AcMNPV p6.9 promoter which provides for earlier expression than the polh promoter. All pOET transfer vectors have a Col E1 origin of replication and an ampicillin resistance gene for selection in E. coli.

The vectors are smaller than other available transfer vectors and range in size between ca 4,500bp and ca 4,700bp, which greatly facilitates the cloning process. The polh coding sequences have been replaced by a multiple cloning site (MCS) containing multiple unique restriction sites for insertion of the foreign gene. The AcMNPV sequences flanking the MCS facilitate recombination with the virus DNA to insert the expression cassette into the polh locus. As well as Oxford Expression Technologies flashBAC platform, these vectors can also be used with any other baculovirus system that uses homologous recombination to insert the foreign gene into the virus genome.
 
Common features of ALL pOET Transfer Vectors:
  • Small size simplifies the cloning process
  • High level expression is achieved under the control of powerful promoters (either the polh promoter or the p6.9 promoter)
  • Directional cloning of gene of interest into the vector is made possible by the presence of multiple unique restriction sites
  • Col E1 origin of replication and an ampicillin resistance gene for selection in E. coli
  • AcMNPV sequences flanking the multiple cloning site (MCS) facilitates recombination with the virus DNA to insert the expression cassette into the polh locus
  • Perfect companion product to the flashBAC™ system
  • Compatible with other baculovirus systems that use homologous recombination
Go to Top
Product Product Features
pOET1™ and pOET2™

Go to Oxford Expression Technologies website
pOET1 and pOET2 are baculovirus transfer vectors designed for high level expression of foreign genes under the powerful AcMNPV polyhedrin (polh) promoter. pOET2 has the same MCS as pOET1, but in the reverse orientation.

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 MCS facilitate recombination with the virus DNA to insert the expression cassette into the polh locus.
 
  • Small size (pOET1 - 4,541bp and pOET2 - 4,547bp) which simplifies the cloning process
  • High level expression from the polh promoter
  • Directional cloning into vector possible using unique restriction sites
  • Supports the expression of truncated proteins
  • pOET1 MCS contains 14 unique restriction sites for insertion of the foreign gene in the correct orientation. pOET2 has the same MCS as pOET1, but in the reverse orientation.
Go to Top
pOET3™ and pOET4™

Go to Oxford Expression Technologies website
pOET3 and pOET4 vectors combine the flexibility and convenience afforded by pOET1 and pOET2 with additional enhanced capabilities that complement the improvements in protein expression delivered by the flashBAC vectors.

Protein expression is driven from the AcMNPV p6.9 promoter which provides for earlier expression than the polh promoter used in pOET1 and pOET2. This can deliver significant improvements when proteins that require extensive post-translational modifications are produced, such as glycoproteins destined for secretion or membrane insertion. The pOET4 Vector has the same MCS as pOET3, but in the reverse orientation, so offers increased options when designing a cloning strategy.
 
  • Small size (pOET3 - 4,530bp and pOET4 - 4,536bp) which simplifies the cloning process
  • High level expression from the AcMNPV basic (p6.9) promoter
  • Directional cloning into vector possible using unique restriction sites
  • Enhanced expression of proteins requiring extensive post-tranlational modifications such as glycosylation
  •  pOET3 MCS contains 19 unique restriction sites for insertion of the foreign gene in the correct orientation. pOET4 has the same MCS as pOET3, but in the reverse orientation.
Go to Top
pOET5

Go to Oxford Expression Technologies website
pOET5 is a dual promoter baculovirus transfer vector designed for high level expression of two foreign genes simultaneously under the powerful AcMNPV polyhedrin (polh) promoter and the very late p10 promoter. The polh sequences have been replaced by two multiple cloning sites (MCS) containing unique restriction sites for insertion of the foreign genes in the correct orientation.
 
  • Small size (pOET5 - 4,590bp) - makes cloning easier
  • The promoters are in opposite orientations to minimise recombination
  • High level expression from the polh and p10 promoters
  • Ideal for the expression of multi-subunit proteins
  • Two sets of unique restriction sites - Polh MCS has 8 sites and p10 MCS has 7 sites
Go to Top
pOET1C_6xHis™ and pOET2C_6xHis™

Go to Oxford Expression Technologies website
pOET1C_6xHis and pOET2C_6xHis are baculovirus transfer vectors designed for high level expression of foreign genes under the powerful AcMNPV polyhedrin (polh) promoter. The vector encodes an optional C-terminal 6xHis-Tag® fusion sequence that may be utilised. This greatly eases the purification of the recombinant protein since the 6×His-containing fusion proteins bind with high affinity to Ni-NTA Agarose. The polh sequences have been replaced by a multiple cloning site (MCS) containing unique set of restriction sites for insertion of the foreign gene in the correct orientation..
 
  • Small size (pOET1C - 4,563bp and pOET2C - 4,593bp) - makes the cloning steps easy
  • Both vectors encode for an optional C-terminal 6xHis-Tag® fusion sequence
  • High level expression from the polh promoter
  • Directional cloning into vector is possible using unique restriction sites. pOET1C contains a unique set 18 sites and pOET2C contains a unique set of 19 sites
Go to Top
pOET1N_6xHis™ and pOET2N/C_6xHis™

Go to Oxford Expression Technologies website
pOET1N_6xHis and pOET2N/C_6xHis are baculovirus transfer vectors designed for high level expression of foreign genes under the powerful AcMNPV polyhedrin (polh) promoter.

The pOET1N_6xHis vector encodes for an optional N-terminal 6xHis-Tag® fusion sequence that may be utilised if the insert allows readthrough in the correct reading frame.

The pOET2N/C_6xHis vector encodes an N-terminal 6xHis-Tag® fusion sequence that may be utilised if the insert includes a stop codon. There is also a 6xHis-Tag®for C-terminal fusions where the start codon of the insert can be used to replace the start codon supplied in pOET2N/C_6xHis.

This presence of an N- or C-terminal 6×His-Tag® fusion sequence 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. The PacI site at the end of the MCS provides translational stop codons in all three reading frames for expression of truncated proteins.
 
  • Small size (pOET1N - 4,598bp and pOET2N/C - 4,626bp) - makes the cloning steps easy
    Both vectors ecode for an optional N-terminal 6xHis-Tag® fusion sequence
  • pOET2N/C also ecodes for an optional C-terminal 6xHis-Tag® fusion sequence
  • 6×His-Tag® can be removed by incubating the fusion protein in the presence of the proteinase cleavage enzyme Thrombin
  • High level expression from the polh promoter
  • The PacI site at the end of the MCS provides translational stop codons in all three reading frames for expression of truncated proteins
  • Directional cloning into vector is possible using unique restriction sites. pOET1N contains a unique set of 23 sites and pOET2N/C contains a unique set of 21 sites
Go to Top
 

Website by Websta Works