The general aim of the proposed project is to understand the mechanism by which the specificity of the site-specific recombination systems changes during evolution. The Cre protein and its recombinational target, the lox sites of bacteriophage P1 will serve as an experimental model. We will build a multipurpose experimental system by which we plan to identify specific nucleotides within the loxsites and specific amino acids in the Cre protein, responsible for specificity. To achieve this goal we will carry out systematic mutagenesis of the regions suspected to contribute to specificity, both in the sites and in the recombinase and will measure their recombinational properties. Based on this information we intend to change the specificity of the Cre/lox system by combining mutually compatible mutations in the lox sites and in the protein. If successful, this will answer the question how well we understood the evolutionary mechanisms. As a practical outcome, it will provide new varieties of the Cre/lox system with different specificities, increasing its utility to manipulate eukaryotic cells for research purposes.

Agency
National Institute of Health (NIH)
Institute
Fogarty International Center (FIC)
Type
Small Research Grants (R03)
Project #
1R03TW001335-01A1
Application #
6335396
Study Section
International and Cooperative Projects 1 Study Section (ICP)
Program Officer
Katz, Flora N
Project Start
2001-05-01
Project End
2004-04-30
Budget Start
2001-05-01
Budget End
2002-04-30
Support Year
1
Fiscal Year
2001
Total Cost
$39,670
Indirect Cost
Name
Brown University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
001785542
City
Providence
State
RI
Country
United States
Zip Code
02912