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Cloning, sequencing and analysis of spirotetronate biosynthetic gene clusters

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Kourdi Zerikly, Malek (2011) Cloning, sequencing and analysis of spirotetronate biosynthetic gene clusters. PhD thesis, University of Warwick.

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Official URL: http://webcat.warwick.ac.uk/record=b2533335~S1

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Abstract

Spirotetronate antibiotics are natural products that have been shown to have a broad
spectrum of biological activities, including antiviral, antibacterial, antitumor and
antimalarial activities. Two representatives of this group are quartromicins and
chrolactomycin, which are produced by Amycolatopsis orientalis and Streptomyces sp.
569N-3 respectively.
Based on other related natural products with known biosynthetic pathways, the
biosynthetic routes for quartromicins and chrolactomycin were proposed and several
strategies for locating the putative gene clusters directing the biosynthesis of these natural
products were used.
An FkbH-like protein was proposed to be specifically involved in the biosynthetic
incorporation of a glycerol-derived three-carbon unit into the tetronate moieties of
quartromicins and chrolactomycin. It was selected as a target to locate the biosynthetic
gene clusters of tetronate natural products. After aligning sequences of known FkbH-like
proteins, a set of degenerate oligonucleotide primers for amplification of fkbH-like genes
was designed and tested. Fragments of fkbH-like sequences were amplified from several
available strains and sequenced.
Several genomic libraries were created and screened by PCR using the degenerate fkbH
primers. After validating the library, clones containing the putative biosynthetic gene
cluster were identified and sequenced. The obtained sequence data was then assembled
and analysed. Coding sequences were identified, protein functions assigned and a
biosynthetic route was proposed for the biosynthesis of quartromicins and
chrolactomycin.

Item Type: Thesis (PhD)
Subjects: Q Science > QD Chemistry
Library of Congress Subject Headings (LCSH): Antibiotics -- Synthesis, Streptomyces -- Genetics, Biosynthesis
Official Date: March 2011
Dates:
DateEvent
March 2011Submitted
Institution: University of Warwick
Theses Department: Department of Chemistry
Thesis Type: PhD
Publication Status: Unpublished
Supervisor(s)/Advisor: Challis, Gregory L.
Extent: xiv, 149 leaves : ill.
Language: eng

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