Regulatory mutants which affect attenuation of the threonine (thr) operon will be isolated and studied. Recently developed in vitro mutagenesis techniques will be employed to direct mutations into specific sites in the thr regulatory region. Mutants will be characterized both by DNA sequencing and by their regulatory effects in vivo. The positions of the mutations and their effects on regulation will supply information on the relationship between secondary RNA structure and transcription termination. Another aspect of the proposal is concerned with generating mutations in the recombination site (att site) used by bacteriophage Lambda during site-specific recombination. Mutations will be induced in vitro and characterized by their effects on recombination in vivo and by their interactions with Lambda integrase (int) protein in DNA protection and binding assays.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM028717-06
Application #
3275997
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1980-12-01
Project End
1987-11-30
Budget Start
1985-12-01
Budget End
1986-11-30
Support Year
6
Fiscal Year
1986
Total Cost
Indirect Cost
Name
University of Illinois Urbana-Champaign
Department
Type
Schools of Arts and Sciences
DUNS #
041544081
City
Champaign
State
IL
Country
United States
Zip Code
61820
Kolakowski, Adam J; Gardner, Jeffrey F (2016) The N-terminus of IntDOT forms hydrophobic interactions during Holliday Junction resolution. Plasmid 87-88:10-16
Wood, Margaret M; Gardner, Jeffrey F (2015) The Integration and Excision of CTnDOT. Microbiol Spectr 3:MDNA3-0020-2014
Ringwald, Kenneth; Gardner, Jeffrey (2015) The Bacteroides thetaiotaomicron protein Bacteroides host factor A participates in integration of the integrative conjugative element CTnDOT into the chromosome. J Bacteriol 197:1339-49
Hopp, Crystal M; Gardner, Jeffrey F; Salyers, Abigail A (2015) The Xis2d protein of CTnDOT binds to the intergenic region between the mob and tra operons. Plasmid 81:63-71
Keeton, Carolyn M; Park, Jiyeon; Wang, Gui-Rong et al. (2013) The excision proteins of CTnDOT positively regulate the transfer operon. Plasmid 69:172-9
Laprise, Jennifer; Yoneji, Sumiko; Gardner, Jeffrey F (2013) IntDOT interactions with core sites during integrative recombination. J Bacteriol 195:1883-91
Keeton, Carolyn M; Hopp, Crystal M; Yoneji, Sumiko et al. (2013) Interactions of the excision proteins of CTnDOT in the attR intasome. Plasmid 70:190-200
Keeton, Carolyn M; Gardner, Jeffrey F (2012) Roles of Exc protein and DNA homology in the CTnDOT excision reaction. J Bacteriol 194:3368-76
Kim, Seyeun; Gardner, Jeffrey F (2011) Resolution of Holliday junction recombination intermediates by wild-type and mutant IntDOT proteins. J Bacteriol 193:1351-8
Kim, Seyeun; Swalla, Brian M; Gardner, Jeffrey F (2010) Structure-function analysis of IntDOT. J Bacteriol 192:575-86

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