Expression of the blue copper protein azurin from Pseudomonas aeruginosa in Escherichia coli

The structural gene for the blue copper protein azurin from Pseudomonas aeruginosa has been subcloned in different expression plasmid vectors. The highest yield of expression was obtained when the gene with its native ribosome-binding site was placed downstream of the lac promoter in plasmid pUC18....

Full description

Saved in:
Bibliographic Details
Published in:FEBS letters 1989-03, Vol.246 (1), p.211-217
Main Authors: Karlsson, B.Göran, Pascher, Torbjörn, Nordling, Margareta, Arvidsson, Rolf H.A., Lundberg, Lennart G.
Format: Article
Language:English
Subjects:
Ap, ampicillin
Azurin
Azurin - genetics
Azurin - isolation & purification
Bacterial Proteins - genetics
Binding Sites
BSA, bovine serum albumin
Cloning
Cloning, Molecular
DNA Restriction Enzymes
Escherichia coli
Escherichia coli - genetics
Expression
gene expression
Gene Expression Regulation
Genes
Genes, Bacterial
Genetic Vectors
IPTG, isopropyl-β-D-thiogalactopyranoside
Nucleic Acid Hybridization
PAGE, polyacrylamide gel electrophoresis
PBS, phosphate-buffered saline
PEG, polyethylene glycol
Plasmids
Promoter Regions, Genetic
Pseudomonas aeruginosa - genetics
Purification
Ribosomes - metabolism
Transformation, Bacterial
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The structural gene for the blue copper protein azurin from Pseudomonas aeruginosa has been subcloned in different expression plasmid vectors. The highest yield of expression was obtained when the gene with its native ribosome-binding site was placed downstream of the lac promoter in plasmid pUC18. The protein is exported to the periplasmic space in Escherichia coli and the amount corresponds to 27% of the total protein content in the periplasmic space. The preprotein is cleaved correctly according to N-terminal sequencing of the purified protein. Azurin has been purified in large amounts and is spectroscopically indistinguishable from the protein purified from P. aeruginosa.
ISSN:0014-5793
1873-3468
DOI:10.1016/0014-5793(89)80285-6