Abstract

Escherichia coli is the best known cellular organism, but many of its genes remain to be identified. The experiments proposed in this revised application will identify previously unknown genes essential for E. coli viability, and also regions of the bacterial chromosome that are free of essential genes, using transposon-based reverse genetic strategies. We will, in particular, identify essential genes that: (i) are expressed only at low levels, (ii) are needed for growth only aerobically, only anaerobically, or only at particular temperatures; and (iii) that encode membrane proteins. Some of these genes will be sequenced, and their protein products characterized biochemically and genetically. Their roles in bacterial growth will be deduced using conditional (heat- and cold- sensitive) mutations. These studies will characterize genes whose products could serve as good targets for the rational design of new drugs of importance in microbial infections, and should provide significant new insights into fundamental principles of bacterial growth.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Human Genome Research Institute (NHGRI)
Type
Research Project (R01)
Project #
5R01HG000563-02
Application #
3333742
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1991-04-01
Project End
1994-03-31
Budget Start
1992-04-01
Budget End
1993-03-31
Support Year
2
Fiscal Year
1992
Total Cost
Indirect Cost

  Institution

Name
Washington University
Department
Type
Schools of Medicine
DUNS #
062761671
City
Saint Louis
State
MO
Country
United States
Zip Code
63130

  Publications

Brikun, I; Suziedelis, K; Stemmann, O et al. (1996) Analysis of CRP-CytR interactions at the Escherichia coli udp promoter. J Bacteriol 178:1614-22
Krishnan, B R; Jamry, I; Berg, D E et al. (1995) Construction of a genomic DNA 'feature map' by sequencing from nested deletions: application to the HLA class I region. Nucleic Acids Res 23:117-22
Akopyants, N S; Eaton, K A; Berg, D E (1995) Adaptive mutation and cocolonization during Helicobacter pylori infection of gnotobiotic piglets. Infect Immun 63:116-21
Kersulyte, D; Struelens, M J; Deplano, A et al. (1995) Comparison of arbitrarily primed PCR and macrorestriction (pulsed-field gel electrophoresis) typing of Pseudomonas aeruginosa strains from cystic fibrosis patients. J Clin Microbiol 33:2216-9
Persson, B C; Bylund, G O; Berg, D E et al. (1995) Functional analysis of the ffh-trmD region of the Escherichia coli chromosome by using reverse genetics. J Bacteriol 177:5554-60
Bukanov, N; Ravi, V N; Miller, D et al. (1994) Pseudomonas aeruginosa corneal ulcer isolates distinguished using the arbitrarily primed PCR DNA fingerprinting method. Curr Eye Res 13:783-90
Woods, J P; Kersulyte, D; Tolan Jr, R W et al. (1994) Use of arbitrarily primed polymerase chain reaction analysis to type disease and carrier strains of Neisseria meningitidis isolated during a university outbreak. J Infect Dis 169:1384-9
Wang, G; Xu, X; Chen, J M et al. (1994) Inversions and deletions generated by a mini-gamma delta (Tn1000) transposon. J Bacteriol 176:1332-8
Bukanov, N O; Berg, D E (1994) Ordered cosmid library and high-resolution physical-genetic map of Helicobacter pylori strain NCTC11638. Mol Microbiol 11:509-23
Fonstein, M; Nikolskaya, T; Zaporojets, D et al. (1994) Tn10-mediated inversions fuse uridine phosphorylase (udp) and rRNA genes of Escherichia coli. J Bacteriol 176:2265-71

Showing the most recent 10 out of 27 publications