Optimization of conditions required for the characterization of Escherichia coli promoters responsive to inter-species interaction with commensal partners : Klebsiella pneumoniae and Enterococcus faecalis.
A promoter-capture library (Lib39) was previously constructed in Escherichia coli based on the dual fluorescence system, which can distinguish E. coli from other species and monitor promoter activities. By using Lib39, a strategy was designed to determine the effect of Klebsiella pneumoniae and Ente...
Saved in:
Main Author: | |
---|---|
Other Authors: | |
Format: | Final Year Project |
Language: | English |
Published: |
2009
|
Subjects: | |
Online Access: | http://hdl.handle.net/10356/16341 |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
Institution: | Nanyang Technological University |
Language: | English |
id |
sg-ntu-dr.10356-16341 |
---|---|
record_format |
dspace |
spelling |
sg-ntu-dr.10356-163412023-02-28T18:04:11Z Optimization of conditions required for the characterization of Escherichia coli promoters responsive to inter-species interaction with commensal partners : Klebsiella pneumoniae and Enterococcus faecalis. Kong, Kiat Whye. Sze Chun Chau School of Biological Sciences DRNTU::Science::Biological sciences::Microbiology::Bacteria A promoter-capture library (Lib39) was previously constructed in Escherichia coli based on the dual fluorescence system, which can distinguish E. coli from other species and monitor promoter activities. By using Lib39, a strategy was designed to determine the effect of Klebsiella pneumoniae and Enterococcus faecalis on E. coli promoter activities. This study aims to verify the feasibility of this strategy. FSC-M/SSC-M subpopulation of K. pneumonia and FSC-H and SSC-H subpopulations of E. faecalis are preferred after analyzing their recovery percentage. After sequencing 40 randomly picked library clones, Lib39 was found to have a good coverage of E. coli genome and promoters. FACSAriaTM was unable to detect the assumed red fluorescent (R+) percentage, which may be due to the limitation of FACSAriaTM in detecting R+ events. Culturing in various temperatures and incubation times were unable to increase the library’s R+ events. A clone, containing a constitutive promoter upstream of mAsRed2, showed an increase in red intensity after static incubation in various conditions, implying that mAsRed2 is functioning but requires longer time to mature. Presence of A-U rich region in the untranslated mRNA may be the reason for ineffective mAsRed2 expression in some of the promoter-mAsRed2 fusions due to RNase E degradation. Bachelor of Science in Biological Sciences 2009-05-25T06:43:05Z 2009-05-25T06:43:05Z 2009 2009 Final Year Project (FYP) http://hdl.handle.net/10356/16341 en Nanyang Technological University 41 p. application/pdf |
institution |
Nanyang Technological University |
building |
NTU Library |
continent |
Asia |
country |
Singapore Singapore |
content_provider |
NTU Library |
collection |
DR-NTU |
language |
English |
topic |
DRNTU::Science::Biological sciences::Microbiology::Bacteria |
spellingShingle |
DRNTU::Science::Biological sciences::Microbiology::Bacteria Kong, Kiat Whye. Optimization of conditions required for the characterization of Escherichia coli promoters responsive to inter-species interaction with commensal partners : Klebsiella pneumoniae and Enterococcus faecalis. |
description |
A promoter-capture library (Lib39) was previously constructed in Escherichia coli based on the dual fluorescence system, which can distinguish E. coli from other species and monitor promoter activities. By using Lib39, a strategy was designed to determine the effect of Klebsiella pneumoniae and Enterococcus faecalis on E. coli promoter activities. This study aims to verify the feasibility of this strategy. FSC-M/SSC-M subpopulation of K. pneumonia and FSC-H and SSC-H subpopulations of E. faecalis are preferred after analyzing their recovery percentage. After sequencing 40 randomly picked library clones, Lib39 was found to have a good coverage of E. coli genome and promoters. FACSAriaTM was unable to detect the assumed red fluorescent (R+) percentage, which may be due to the limitation of FACSAriaTM in detecting R+ events. Culturing in various temperatures and incubation times were unable to increase the library’s R+ events. A clone, containing a constitutive promoter upstream of mAsRed2, showed an increase in red intensity after static incubation in various conditions, implying that mAsRed2 is functioning but requires longer time to mature. Presence of A-U rich region in the untranslated mRNA may be the reason for ineffective mAsRed2 expression in some of the promoter-mAsRed2 fusions due to RNase E degradation. |
author2 |
Sze Chun Chau |
author_facet |
Sze Chun Chau Kong, Kiat Whye. |
format |
Final Year Project |
author |
Kong, Kiat Whye. |
author_sort |
Kong, Kiat Whye. |
title |
Optimization of conditions required for the characterization of Escherichia coli promoters responsive to inter-species interaction with commensal partners : Klebsiella pneumoniae and Enterococcus faecalis. |
title_short |
Optimization of conditions required for the characterization of Escherichia coli promoters responsive to inter-species interaction with commensal partners : Klebsiella pneumoniae and Enterococcus faecalis. |
title_full |
Optimization of conditions required for the characterization of Escherichia coli promoters responsive to inter-species interaction with commensal partners : Klebsiella pneumoniae and Enterococcus faecalis. |
title_fullStr |
Optimization of conditions required for the characterization of Escherichia coli promoters responsive to inter-species interaction with commensal partners : Klebsiella pneumoniae and Enterococcus faecalis. |
title_full_unstemmed |
Optimization of conditions required for the characterization of Escherichia coli promoters responsive to inter-species interaction with commensal partners : Klebsiella pneumoniae and Enterococcus faecalis. |
title_sort |
optimization of conditions required for the characterization of escherichia coli promoters responsive to inter-species interaction with commensal partners : klebsiella pneumoniae and enterococcus faecalis. |
publishDate |
2009 |
url |
http://hdl.handle.net/10356/16341 |
_version_ |
1759854237721821184 |