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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 BJMBR
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
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Genome features of Leptospira interrogans serovar Copenhageni BJMBR
Nascimento,A.L.T.O.; Verjovski-Almeida,S.; Van Sluys,M.A.; Monteiro-Vitorello,C.B.; Camargo,L.E.A.; Digiampietri,L.A.; Harstkeerl,R.A.; Ho,P.L.; Marques,M.V.; Oliveira,M.C.; Setubal,J.C.; Haake,D.A.; Martins,E.A.L..
We report novel features of the genome sequence of Leptospira interrogans serovar Copenhageni, a highly invasive spirochete. Leptospira species colonize a significant proportion of rodent populations worldwide and produce life-threatening infections in mammals. Genomic sequence analysis reveals the presence of a competent transport system with 13 families of genes encoding for major transporters including a three-member component efflux system compatible with the long-term survival of this organism. The leptospiral genome contains a broad array of genes encoding regulatory system, signal transduction and methyl-accepting chemotaxis proteins, reflecting the organism's ability to respond to diverse environmental stimuli. The identification of a complete set...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Leptospira interrogans; Outer membrane proteins; Lipopolysaccharides; Transport; Regulatory systems.
Ano: 2004 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2004000400003
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