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Analysis of secondary metabolite gene and protein expression profiles in Streptomyces coelicolor grown under environmental conditions. / Kathryn Laura Bell

Swansea University Author: Kathryn Laura Bell

Abstract

Streptomycetes are Gram positive, soil dwelling filamentous bacteria, known for their production of secondary metabolites. Genome sequencing of Streptomyces coelicolor identified 26 known or predicted secondary metabolite gene clusters ranging from antibiotics to siderophores, lipids, pigments and l...

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Published: 2012
Institution: Swansea University
Degree level: Doctoral
Degree name: Ph.D
URI: https://cronfa.swan.ac.uk/Record/cronfa42693
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last_indexed 2019-10-21T16:48:17Z
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spelling 2018-08-29T14:29:29.4346979 v2 42693 2018-08-02 Analysis of secondary metabolite gene and protein expression profiles in Streptomyces coelicolor grown under environmental conditions. 0662954d40a3b3c2d92cba3863752c80 NULL Kathryn Laura Bell Kathryn Laura Bell true true 2018-08-02 Streptomycetes are Gram positive, soil dwelling filamentous bacteria, known for their production of secondary metabolites. Genome sequencing of Streptomyces coelicolor identified 26 known or predicted secondary metabolite gene clusters ranging from antibiotics to siderophores, lipids, pigments and lantibiotics. Most studies investigating secondary metabolite production in Streptomyces, as well as other bacteria, are undertaken in liquid or on solid media. There is little gene expression data available from in situ studies. This study determined expression in a totally different growth medium, soil, to gain insight into growth and adaptation under more 'normal' habitat conditions and the effect changing environmental factors has on the expression of secondary metabolite gene clusters. In order to do this, S. coelicolor was grown in soil microcosms from which RNA was extracted and amplified using an optimised T7 polymerase-based RNA amplification protocol. The amplified RNA was used to determine gene expression profiles via endpoint RT-PCR and RT-qPCR. In a complementary approach, S. coelicolor sand microcosms were subjected to a novel protein extraction procedure to determine protein expression profiles in soil. This study elucidated how carbon, nitrogen and metal availability, the environmentally bioactive entomopathogenic fungus Metarhizium anisopliae and nematode Steinernema kraussei affect secondary metabolite gene expression in soil. In contrast to the consensus of secondary metabolism commencing after reduction or cessation of growth, this study revealed expression of secondary metabolite biosynthetic genes and proteins related to secondary metabolism before the onset of exponential growth. Some secondary metabolite genes/proteins were even expressed constitutively. Soil microcosms have been shown to be an important tool for gene expression analysis. The results of these novel transcriptomic and proteomic approaches therefore have given new insight into secondary metabolism and its role under natural habitat conditions. E-Thesis Genetics.;Microbiology. 31 12 2012 2012-12-31 COLLEGE NANME Swansea University Medical School COLLEGE CODE Swansea University Doctoral Ph.D 2018-08-29T14:29:29.4346979 2018-08-02T16:24:30.1334020 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Medicine Kathryn Laura Bell NULL 1 0042693-02082018162514.pdf 10807462.pdf 2018-08-02T16:25:14.5300000 Output 18257769 application/pdf E-Thesis true 2018-08-02T16:25:14.5300000 false
title Analysis of secondary metabolite gene and protein expression profiles in Streptomyces coelicolor grown under environmental conditions.
spellingShingle Analysis of secondary metabolite gene and protein expression profiles in Streptomyces coelicolor grown under environmental conditions.
Kathryn Laura Bell
title_short Analysis of secondary metabolite gene and protein expression profiles in Streptomyces coelicolor grown under environmental conditions.
title_full Analysis of secondary metabolite gene and protein expression profiles in Streptomyces coelicolor grown under environmental conditions.
title_fullStr Analysis of secondary metabolite gene and protein expression profiles in Streptomyces coelicolor grown under environmental conditions.
title_full_unstemmed Analysis of secondary metabolite gene and protein expression profiles in Streptomyces coelicolor grown under environmental conditions.
title_sort Analysis of secondary metabolite gene and protein expression profiles in Streptomyces coelicolor grown under environmental conditions.
author_id_str_mv 0662954d40a3b3c2d92cba3863752c80
author_id_fullname_str_mv 0662954d40a3b3c2d92cba3863752c80_***_Kathryn Laura Bell
author Kathryn Laura Bell
author2 Kathryn Laura Bell
format E-Thesis
publishDate 2012
institution Swansea University
college_str Faculty of Medicine, Health and Life Sciences
hierarchytype
hierarchy_top_id facultyofmedicinehealthandlifesciences
hierarchy_top_title Faculty of Medicine, Health and Life Sciences
hierarchy_parent_id facultyofmedicinehealthandlifesciences
hierarchy_parent_title Faculty of Medicine, Health and Life Sciences
department_str Swansea University Medical School - Medicine{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Medicine
document_store_str 1
active_str 0
description Streptomycetes are Gram positive, soil dwelling filamentous bacteria, known for their production of secondary metabolites. Genome sequencing of Streptomyces coelicolor identified 26 known or predicted secondary metabolite gene clusters ranging from antibiotics to siderophores, lipids, pigments and lantibiotics. Most studies investigating secondary metabolite production in Streptomyces, as well as other bacteria, are undertaken in liquid or on solid media. There is little gene expression data available from in situ studies. This study determined expression in a totally different growth medium, soil, to gain insight into growth and adaptation under more 'normal' habitat conditions and the effect changing environmental factors has on the expression of secondary metabolite gene clusters. In order to do this, S. coelicolor was grown in soil microcosms from which RNA was extracted and amplified using an optimised T7 polymerase-based RNA amplification protocol. The amplified RNA was used to determine gene expression profiles via endpoint RT-PCR and RT-qPCR. In a complementary approach, S. coelicolor sand microcosms were subjected to a novel protein extraction procedure to determine protein expression profiles in soil. This study elucidated how carbon, nitrogen and metal availability, the environmentally bioactive entomopathogenic fungus Metarhizium anisopliae and nematode Steinernema kraussei affect secondary metabolite gene expression in soil. In contrast to the consensus of secondary metabolism commencing after reduction or cessation of growth, this study revealed expression of secondary metabolite biosynthetic genes and proteins related to secondary metabolism before the onset of exponential growth. Some secondary metabolite genes/proteins were even expressed constitutively. Soil microcosms have been shown to be an important tool for gene expression analysis. The results of these novel transcriptomic and proteomic approaches therefore have given new insight into secondary metabolism and its role under natural habitat conditions.
published_date 2012-12-31T03:53:28Z
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score 11.036706

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