Consulta: creatorFacets:"Abreu,P.A.E."
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A high-copy T7 Escherichia coli expression vector for the production of recombinant proteins with a minimal N-terminal His-tagged fusion peptide
Provedor de dados: 56
Autores: Ramos,C.R.R.; Abreu,P.A.E.; Nascimento,A.L.T.O.; Ho,P.L.
We report here the construction of a vector derived from pET3-His and pRSET plasmids for the expression and purification of recombinant proteins in Escherichia coli based on T7 phage RNA polymerase. The resulting pAE plasmid combined the advantages of both vectors: small size (pRSET), expression of a short 6XHis tag at N-terminus (pET3-His) and a high copy number of plasmid (pRSET). The small size of the vector (2.8 kb) and the high copy number/cell (200-250 copies) facilitate the subcloning and sequencing procedures when compared to the pET system (pET3-His, 4.6 kb and 40-50 copies) and also result in high level expression of recombinant proteins (20 mg purified protein/liter of culture). In addition, the vector pAE enables the expression of a fusion...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Escherichia coli; Expression vector; Immobilized metal affinity chromatography; Protein purification.
Ano: 2004
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2004000800001