Mutational Analysis of NBU1 Attachment Sites Reveals a Novel Recombination Mechanism
Schmidt, John William
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https://hdl.handle.net/2142/86675
Description
Title
Mutational Analysis of NBU1 Attachment Sites Reveals a Novel Recombination Mechanism
Author(s)
Schmidt, John William
Issue Date
2004
Doctoral Committee Chair(s)
Gardner, Jeffrey F.
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, Microbiology
Language
eng
Abstract
NBU1 is a 10.3 kbp mobilizable transposon that is normally integrated in the chromosome of its Bacteroides host. Bacteroides are the predominant member of the human intestinal tract microflora, and are opportunistic pathogens that can cause life-threatening infections. A recent survey of Bacteroides isolates has shown an increase in tetracycline resistance, which has been attributed to mobilizable and related conjugative transposons. NBU1 integrates site specifically into the Bacteroides thetaiotaomicron chromosome primary target site, attBT1-1. NBU1 integrase (IntN1) catalyzes recombination of the NBU1 attachment site, attN1, and the attBT1-1 site. IntN1 is a member of the lambda family of tyrosine site-specific recombinases. However, the mechanisms of integration of NBU1 and related mobilizable transposons have not been fully characterized and may be unique. Previous studies have shown that NBU1 has a broad host range and can integrate nonspecifically in Escherichia coli. An in vivo E. coli integration system has been developed to study the nucleotide requirements for NBU1 site-specific integration. This in vivo system has been used to determine the minimal attN1 site. Mutational analysis of the attN1 site has indicated that several nucleotide sequence motifs in the attN1 site are required for integration. Analysis of attN1 and attBT1-1 mutations has also led to the conclusion that recombination mechanism of NBU1 is novel.
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