This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Sankar, P Articles by Neidhardt, F C Search for Related Content PubMed PubMed Citation Articles by Sankar, P Articles by Neidhardt, F C

research-article

Expression analysis of cloned chromosomal segments of Escherichia coli.

Department of Microbiology and Immunology, University of Michigan, Ann Arbor 48109-0620.

ABSTRACT

The novel transcription system of bacteriophage T7 was used to express Escherichia coli genes preferentially with a new low-copy-number plasmid vector, pFN476, to minimize toxic gene effects. Selected E. coli chromosomal fragments from an ordered genomic library (Y. Kohara, K. Ikiyama, and K. Isono, Cell 50:495-508, 1987) were recloned into this vector, and their genes were preferentially expressed in vivo utilizing its T7 promoter. The protein products were analyzed by two-dimensional gel electrophoresis. By using DNA sequence information, the gel migration was predicted for the protein products of open reading frames from these segments, and this information was used to identify gene products visualized as spots on two-dimensional gels. Even in the absence of DNA sequence information, this approach offers the opportunity to identify all gene products of E. coli and map their genes to within 10 kb on the E. coli genome; with sequence information, this approach can produce a definitive expression map of the E. coli genome.

This article has been cited by other articles:

• Han, M.-J., Lee, S. Y. (2006). The Escherichia coli Proteome: Past, Present, and Future Prospects. Microbiol. Mol. Biol. Rev. 70: 362-439 [Abstract] [Full Text]  
• Huang, S.-H., Chen, Y.-H., Fu, Q., Stins, M., Wang, Y., Wass, C., Kim, K. S. (1999). Identification and Characterization of an Escherichia coli Invasion Gene Locus, ibeB, Required for Penetration of Brain Microvascular Endothelial Cells. Infect. Immun. 67: 2103-2109 [Abstract] [Full Text]  
• Chatterjee, S., Mondal, A. K., Begum, N. A., Roychoudhury, S., Das, J. (1998). Ordered Cloned DNA Map of the Genome of Vibrio cholerae 569B and Localization of Genetic Markers. J. Bacteriol. 180: 901-908 [Abstract] [Full Text]  
• Davis, M. S., Solbiati, J., Cronan, J. E. Jr. (2000). Overproduction of Acetyl-CoA Carboxylase Activity Increases the Rate of Fatty Acid Biosynthesis in Escherichia coli. J. Biol. Chem. 275: 28593-28598 [Abstract] [Full Text]