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|Title:||Bacteroides Conjugative Transposons: Ecology and Mechanisms|
|Doctoral Committee Chair(s):||Abigail Salyers|
|Department / Program:||Microbiology|
|Degree Granting Institution:||University of Illinois at Urbana-Champaign|
|Abstract:||CTnDOT is a 65 kb Bacteroides CTn that carries genes encoding for tetracycline resistance and erythromycin resistance. Previous studies had shown that the attDOT recombination site and integrase of CTnDOT (IntDOT) are sufficient and essential for integration in vivo. Even though the integrase of CTnDOT belongs to the lambda integrase family of recombinases, the mechanism of integration of CTnDOT is different from that of phage lambda, the model for site-specific recombination systems. Although IntDOT is sufficient for integration in vivo, recent studies have shown that a host factor is required for efficient integration. Using the gel shift assay technique, we have shown that IntDOT binds attDOT weakly and binding of IntDOT is enhanced in the presence of IHF. We propose that IHF bends attDOT DNA during recombination, facilitating the binding of CTnDOT integrase to attDOT. Using a combination of the in vivo integration assay and the gel shift assay, attDOT has been narrowed to 266 bp. We have identified the sites where IntDOT and IHF bind to attDOT using the footprinting technique.|
Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 2004.
|Date Available in IDEALS:||2015-09-28|