Mammalian Expression System – Xpressª System Protein Expression pEBVHis – Invitrogen

mamalian expression system

The pEBVHis A, B, and C vectors are derived from the Epstein Barr Virus (EBV) expression vectorpREP4. They are designed for efficient protein expression and purification in mammalian tissue-culturedcells.High levels of expression of DNA sequences cloned into the pEBVHis vectors are made possibleby the presence of the EBV origin of replication, designated ori-P, and the EBNA-1 protein (EBV-encoded nuclear antigen) which trans-activates ori-P. The inclusion of these elements in the EBVvectors enables them to replicate episomally when transfected into an appropriate mammalian cell line(e.g. 293, 293-EBNA, COS, or CV-1). They will not replicate in rodent cell lines (e.g. CHO or NIH-3T3).The vectors can be maintained as high copy number plasmids for up to 3 months, if selective pressure ismaintained, making it no longer necessary to produce a stably transformed clonal derivative.Transcription of the gene of interest is directed by the Rous Sarcoma Virus long terminal repeat(RSV-LTR). The vectors carry the Hygromycin B drug resistance marker under the control of theThymidine Kinase (TK) promoter, enabling selection in mammalian cells with hygromycin. The vector contains a sequence which codes for (in 5′ to 3′ direction from N-terminal to C-terminal)an ATG translation initiation codon, a series of six histidine residues that function as a metal bindingdomain in the translated protein, a transcript stabilizing sequence from gene 10 of phage T7, and anenterokinase cleavage recognition sequence. A multiple cloning site positioned downstream of thissequence allows insertion of the foreign gene in the correct reading frame relative to the initiation codon.The metal binding domain of the fusion peptide allows simple one step purification of recombinantproteins by Immobilized Metal Affinity Chromatography, which is made possible with Invitrogen’sProBond™ resin (available as prepackaged disposable columns or in bulk). The enterokinase cleavagerecognition site in the fusion peptide between the metal binding domain and the recombinant proteinallows for subsequent removal of this N-terminal fusion peptide from the purified recombinant protein

Invitrogen mamalian expression system


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