Bacteroides Conjugative Transposons: Ecology and Mechanisms

Gupta, Anamika

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Description

Title

Bacteroides Conjugative Transposons: Ecology and Mechanisms

Author(s)

Gupta, Anamika

Issue Date

2004

Doctoral Committee Chair(s)

Abigail Salyers

Department of Study

Microbiology

Discipline

Microbiology

Degree Granting Institution

University of Illinois at Urbana-Champaign

Degree Name

Ph.D.

Degree Level

Dissertation

Keyword(s)

Biology, Molecular

Language

eng

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.

Graduation Semester

2004

Type of Resource

text

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