Abstract

Klebsiella pneumoniae is an enteric opportunistic pathogen which commonly causes pulmonary, urinary tract and kidney infections. It has also been widely used for the study of nitrogen fixation. In this study, a macrorestriction map of K. pneumoniae M5a1 chromosomal DNA was constructed using pulsed field gel electrophoresis (PFGE) and Southern hybridization techniques. The genome DNA was digested with two rare cutting restriction enzymes (XbaI and BlnI) and the genome size was estimated from the sum of the fragment sizes as 5,121 Kb (+2.8%). A tentative order of fragments was deduced using Southern hybridization with purified restriction fragments. Several auxotrophic mutants were isolated from M5a1 and the chromosomal clones that complemented them were identified. Random clones from the M5a1 chromosomal library were also obtained. These clones and clones acquired from other laboratories were mapped onto the circular macrorestriction map of M5a1. The order of fragments obtained by this means agreed with the map constructed by Southern hybridization of Blnl and Xbal fragments.

This map can be used to construct a typing system for clinical K. pneumoniae isolates based upon DNA fingerprinting. It will also facilitate the construction of a high resolution (microrestriction) map and the determination of the complete nucleotide sequence of the organism. Preliminary genomic analyses of clinical samples show that there are large variations in the genomic pattern of K. pneumoniae samples from different patients even though they are isolated from the same location of the body.

LLU Discipline

Biology

Department

Biology

School

Graduate School

First Advisor

Junichi Ryu

Second Advisor

Leonard R. Brand

Third Advisor

Ronald L. Carter

Fourth Advisor

David L. Cowles

Fifth Advisor

Joseph G. Galusha

Sixth Advisor

Anthony J. Zuccarelli

Degree Name

Doctor of Philosophy (PhD)

Degree Level

Ph.D.

Year Degree Awarded

1994

Date (Title Page)

3-1994

Language

English

Library of Congress/MESH Subject Headings

Klebsiella pneumoniae -- genetics; Genome, Bacterial; Restriction Mapping

Type

Dissertation

Page Count

2; viii

Digital Format

PDF

Digital Publisher

Loma Linda University Libraries

Usage Rights

This title appears here courtesy of the author, who has granted Loma Linda University a limited, non-exclusive right to make this publication available to the public. The author retains all other copyrights.

Collection

Loma Linda University Electronic Theses and Dissertations

Collection Website

http://scholarsrepository.llu.edu/etd/

Repository

Loma Linda University. Del E. Webb Memorial Library. University Archives

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