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Acquisition of Storm Phosphorimager for Study of Gene Expression

Investigators
Lee, Theodore
Institutions
State University of New York
Start date
2000
End date
2002
Objective
The research will focus on characterizing the regulation of gene expression of the groESL and dnaKJ operons in Clostridium perfringens and Rhodobacter sphaeroides.
More information
A Molecular Dynamics Storm PhosphorImager system will be used for the study of molecular chaperone gene expression and regulation in prokaryotic systems. The Molecular Dynamics Storm system is approximately ten times more sensitive than X-ray film and has 2.5 times the dynamic range. These properties and the data analysis software provided will allow for easy, fast, and accurate quantitation of data. This makes the Storm system an essential tool for looking at low levels of expression and accurate quantitation of labeled oligonucleotides for determining the dissociation constants for protein-DNA binding. The Storm system is amenable to the detection of radioactive isotopes and to chemifluorescence thus providing a non-radioactive option for students doing research while reducing radioactive isotope storage and disposal problems. The research will focus on characterizing the regulation of gene expression of the groESL and dnaKJ operons in Clostridium perfringens and Rhodobacter sphaeroides.

The groESL and dnaKJ operons encode molecular chaperones that are conserved in prokaryotic and eukaryotic organisms and play a critical role in the folding of proteins in cells. In C. perfringens, a food-borne pathogen, the research will combine the study of gene expression with the analysis of regulatory sequences and structural investigations of the protein-DNA complexes involved in regulation. By understanding the regulatory mechanisms of these systems it may be possible to design therapeutic compounds that inhibit the expression of these critical molecular chaperone proteins. The regulation of the molecular chaperone genes in R. sphaeroides is also of interest due to the different regulatory sequences present in the groESL and dnaKJ operons. This differential expression is unusual and may serve an important comparison model to the regulatory mechanisms of other bacteria. The combination of genetic and biochemical approaches to understand the regulatory mechanism of gene expression will complement each other and promote scientific research in both departments.

Funding Source
United States Nat'l. Science Fndn.
Project number
0079465
Categories
Bacterial Pathogens
Clostridium