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.

By M. Kitagawa, Takeshi Ara, Mohammad Arifuzzaman, Tomoko Ioka-Nakamichi, E. Inamoto, Hiromi Toyonaga, Hirotada Mori. DNA Research, 2006

1,445 citations recorded by the source corpus.

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.

Subfields: Ecology, Genetics, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/a4a4d5c761f30090fae6ea9446092edb51f5df53

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In Ecology

1. K. Allison
2. Ahmad S. Khalil
3. James J. Collins
4. Daniel J. Dwyer

In Genetics

1. James J. Collins
2. K. Allison
3. Ahmad S. Khalil
4. V. Schachter
5. Pierre-Yves Bourguignon
6. Maxime Durot
7. Daniel J. Dwyer
8. Dongbin Xiu

In Molecular Biology

1. James J. Collins
2. Ahmad S. Khalil
3. K. Allison
4. Daniel J. Dwyer
5. V. Schachter
6. A. Ting
7. Pierre-Yves Bourguignon
8. Maxime Durot

Citations in the corpus

• Genome-scale models of bacterial metabolism: reconstruction and applications (2008, background): “The availability of large-scale libraries of ORF clones (Kitagawa et al ., 2005) should increase the likelihood of such methods expanding the store of known reactions.”
• Antibiotics induce redox-related physiological alterations as part of their lethality (2014, methodology): “The pCA24N::MutS plasmid for MutS dosage studies was obtained from the ASKA overexpression plasmid library (107).”