Archive clones are a set of plasmid clones containing all predicted ORFs of E. coli. Based on the genomic sequence data of Escherichia coli K-12, strain W3110, a complete set of individual genes encoding Histidine tagged proteins with or without GFP attached was constructed for functional genomic analysis. Each clone encodes a protein of predicted ORF flanked by histidine residues and 7 spacer amino acids at the N terminal end, and 5 spacer amino acids and GFP at the C terminal end. SfiI restriction sites were generated at both the N and C terminal boundaries of each ORF, which enables easy transfer of the ORF into a different vector. Expression of the cloned ORF is under the control of an IPTG-inducible promoter which is strictly repressed by the LacI(gene product of lacIq) repressor. The set of cloned ORFs provides resources for systematic functional genomic approaches including 1) construction of DNA microarray as a DNA source, 2) production and purification of proteins by His tag, 3) analysis of protein localization by monitoring GFP fluorescence, and 4) analysis of protein-protein interactions.
1) Kitagawa, M., Ara, T., Arifuzzaman, M., Ioka-Nakamichi, T., Inamoto, E.,
Toyonaga, H. and Mori, H. Complete set of ORF clones of Escherichia coli
ASKA library (A Complete Set of E. coli K-12 ORF Archive): Unique Resources
for Biological Research. DNA Res 12, 291-299 (2005).
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