Determining the cellular location of FloT and NfeD in streptomyces coelicolor

Erin O’Shaughnessy

Research output: ThesisMaster's Thesis

Abstract

Flotillin proteins are ubiquitous in all kingdoms of life; they have been studied extensively in the model organism Bacillus subtilis and the localisation protein FloT has been described in this model as having a lateral distribution in cells. NfeD is a protein that was first isolated from Sinorhizobium meliloti and was identified as a factor that enhanced nodulation. In B. subtilis these proteins co-localise. We aimed to describe the localisation of these proteins in the filamentous bacteria Streptomyces coelicolor in order to infer functions of these proteins and identify any role that they may play in hyphal growth and development. This study used cloning techniques involving the use of restriction enzymes and ligase to translationally fuse the SCO3607 and SCO3608 genes to mCherry and GFP respectively. We observed the localisation of FloT in samples grown after 24h, 26h, and 72h with the use of fluorescence microscopy. FloT was visualised as GFP foci or diffuse GFP expression within the samples. This research has shown that FloT localises in a random lateral manner in vegetative hyphae in S. coelicolor.
LanguageEnglish
QualificationMPhil
Awarding Institution
  • University Of Strathclyde
Supervisors/Advisors
  • Herron, Paul, Supervisor
Award date2 Jul 2015
Place of PublicationGlasgow
Publisher
Publication statusPublished - 2 Jul 2015
Externally publishedYes

Fingerprint

Streptomyces coelicolor
Proteins
Bacillus subtilis
Sinorhizobium meliloti
Hyphae
Ligases
Growth and Development
Fluorescence Microscopy
Organism Cloning
Bacteria
Enzymes
Research
Genes

Keywords

  • streptomyces coelicolor
  • FloT
  • NfeD

Cite this

O’Shaughnessy, E. (2015). Determining the cellular location of FloT and NfeD in streptomyces coelicolor. Glasgow: University of Strathclyde.
O’Shaughnessy, Erin. / Determining the cellular location of FloT and NfeD in streptomyces coelicolor. Glasgow : University of Strathclyde, 2015.
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title = "Determining the cellular location of FloT and NfeD in streptomyces coelicolor",
abstract = "Flotillin proteins are ubiquitous in all kingdoms of life; they have been studied extensively in the model organism Bacillus subtilis and the localisation protein FloT has been described in this model as having a lateral distribution in cells. NfeD is a protein that was first isolated from Sinorhizobium meliloti and was identified as a factor that enhanced nodulation. In B. subtilis these proteins co-localise. We aimed to describe the localisation of these proteins in the filamentous bacteria Streptomyces coelicolor in order to infer functions of these proteins and identify any role that they may play in hyphal growth and development. This study used cloning techniques involving the use of restriction enzymes and ligase to translationally fuse the SCO3607 and SCO3608 genes to mCherry and GFP respectively. We observed the localisation of FloT in samples grown after 24h, 26h, and 72h with the use of fluorescence microscopy. FloT was visualised as GFP foci or diffuse GFP expression within the samples. This research has shown that FloT localises in a random lateral manner in vegetative hyphae in S. coelicolor.",
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author = "Erin O’Shaughnessy",
year = "2015",
month = "7",
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O’Shaughnessy, E 2015, 'Determining the cellular location of FloT and NfeD in streptomyces coelicolor', MPhil, University Of Strathclyde, Glasgow.

Determining the cellular location of FloT and NfeD in streptomyces coelicolor. / O’Shaughnessy, Erin.

Glasgow : University of Strathclyde, 2015.

Research output: ThesisMaster's Thesis

TY - THES

T1 - Determining the cellular location of FloT and NfeD in streptomyces coelicolor

AU - O’Shaughnessy, Erin

PY - 2015/7/2

Y1 - 2015/7/2

N2 - Flotillin proteins are ubiquitous in all kingdoms of life; they have been studied extensively in the model organism Bacillus subtilis and the localisation protein FloT has been described in this model as having a lateral distribution in cells. NfeD is a protein that was first isolated from Sinorhizobium meliloti and was identified as a factor that enhanced nodulation. In B. subtilis these proteins co-localise. We aimed to describe the localisation of these proteins in the filamentous bacteria Streptomyces coelicolor in order to infer functions of these proteins and identify any role that they may play in hyphal growth and development. This study used cloning techniques involving the use of restriction enzymes and ligase to translationally fuse the SCO3607 and SCO3608 genes to mCherry and GFP respectively. We observed the localisation of FloT in samples grown after 24h, 26h, and 72h with the use of fluorescence microscopy. FloT was visualised as GFP foci or diffuse GFP expression within the samples. This research has shown that FloT localises in a random lateral manner in vegetative hyphae in S. coelicolor.

AB - Flotillin proteins are ubiquitous in all kingdoms of life; they have been studied extensively in the model organism Bacillus subtilis and the localisation protein FloT has been described in this model as having a lateral distribution in cells. NfeD is a protein that was first isolated from Sinorhizobium meliloti and was identified as a factor that enhanced nodulation. In B. subtilis these proteins co-localise. We aimed to describe the localisation of these proteins in the filamentous bacteria Streptomyces coelicolor in order to infer functions of these proteins and identify any role that they may play in hyphal growth and development. This study used cloning techniques involving the use of restriction enzymes and ligase to translationally fuse the SCO3607 and SCO3608 genes to mCherry and GFP respectively. We observed the localisation of FloT in samples grown after 24h, 26h, and 72h with the use of fluorescence microscopy. FloT was visualised as GFP foci or diffuse GFP expression within the samples. This research has shown that FloT localises in a random lateral manner in vegetative hyphae in S. coelicolor.

KW - streptomyces coelicolor

KW - FloT

KW - NfeD

M3 - Master's Thesis

PB - University of Strathclyde

CY - Glasgow

ER -

O’Shaughnessy E. Determining the cellular location of FloT and NfeD in streptomyces coelicolor. Glasgow: University of Strathclyde, 2015.