Extracellular secretion of polypeptides using a modified Escherichia coli flagellar secretion apparatus.
Nat Biotechnol
; 23(4): 475-81, 2005 Apr.
Article
em En
| MEDLINE
| ID: mdl-15806100
ABSTRACT
We developed a modified flagellar type III secretion apparatus to secrete heterologous polypeptides into the growth medium of Escherichia coli. The secretion was facilitated by fusing the 173-bp untranslated DNA fragment upstream of the gene fliC (encoding flagellin) as well as a transcriptional terminator from fliC, into the gene encoding the polypeptide of interest. The polypeptides secreted into the growth medium at concentrations ranging from 1 to 15 mg/l were from Campylobacter jejuni (262 residues in length), Streptococcus pneumoniae (434 residues), Staphylococcus aureus (115 residues), and N-terminal FliC hybrid proteins, for example, the eukaryotic green fluorescent protein (238 residues). The expressed proteins represented >50% of total secreted protein. Previously reported protein yields from extracellular secretion of foreign proteins in E. coli have been low, approximately 100 microg/l. The strengths of our method are the concentration and purity of the secreted proteins and its versatility with regard to the proteins' length and origin.
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Base de dados:
MEDLINE
Assunto principal:
Peptídeos
/
Proteínas de Bactérias
/
Escherichia coli
/
Flagelos
/
Flagelina
Idioma:
En
Ano de publicação:
2005
Tipo de documento:
Article