RRC ID 33728
著者 Kitagawa M, Ara T, Arifuzzaman M, Ioka-Nakamichi T, Inamoto E, Toyonaga H, Mori H.
タイトル 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.
ジャーナル DNA Res
Abstract Based on the genomic sequence data of Escherichia coli K-12 strain, we have constructed a complete set of cloned individual genes encoding Histidine-tagged proteins with or without GFP fused for functional genomic analysis. Each clone encodes a protein of predicted ORF attached by Histidines and seven spacer amino acids at the N-terminal end, and five spacer amino acids and GFP at the C-terminal end. SfiI restriction sites are generated at both the N- and C-terminal boundaries of ORF upon cloning, which enables easy transfer of ORF to other vector systems by cutting with SfiI. Expression of cloned ORF is under the control of an IPTG-inducible promoter, which is strictly repressed by lacI(q) repressor gene product. The set of cloned ORFs described here should provide unique resources for systematic functional genomic approaches including (i) construction of DNA microarray, (ii) production and purification of proteins, (iii) analysis of protein localization by monitoring GFP fluorescence and (iv) analysis of protein-protein interaction.
巻・号 12(5)
ページ 291-9
公開日 2005-1-1
DOI 10.1093/dnares/dsi012
PII dsi012
PMID 16769691
MeSH Escherichia coli K12 / genetics* Escherichia coli K12 / metabolism Escherichia coli Proteins / genetics* Escherichia coli Proteins / metabolism Gene Library* Green Fluorescent Proteins / genetics Green Fluorescent Proteins / metabolism Histidine / chemistry Open Reading Frames* Promoter Regions, Genetic Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism
IF 4.009
引用数 870
WOS 分野 GENETICS & HEREDITY
リソース情報
原核生物(大腸菌) ASKA Clone