PMID:15806100
| Citation |
Majander, K, Anton, L, Antikainen, J, Lång, H, Brummer, M, Korhonen, TK and Westerlund-Wikström, B (2005) Extracellular secretion of polypeptides using a modified Escherichia coli flagellar secretion apparatus. Nat. Biotechnol. 23:475-81 |
|---|---|
| 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. |
| Links |
PubMed Online version:10.1038/nbt1077 |
| Keywords |
Bacterial Proteins/genetics; Bacterial Proteins/secretion; Campylobacter jejuni/chemistry; Culture Media/chemistry; DNA, Bacterial; DNA, Recombinant; Escherichia coli/cytology; Escherichia coli/genetics; Escherichia coli/growth & development; Escherichia coli/metabolism; Flagella/physiology; Flagella/secretion; Flagellin/chemistry; Flagellin/genetics; Flagellin/metabolism; Gene Deletion; Genes, Bacterial; Green Fluorescent Proteins/chemistry; Molecular Sequence Data; Mutagenesis, Site-Directed; Peptides/secretion; Protein Binding; Recombinant Proteins/secretion; Staphylococcus aureus/chemistry; Streptococcus pneumoniae/chemistry |
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