Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/16341
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.
Authors: Kong, Kiat Whye.
Keywords: DRNTU::Science::Biological sciences::Microbiology::Bacteria
Issue Date: 2009
Abstract: 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.
URI: http://hdl.handle.net/10356/16341
Rights: Nanyang Technological University
Fulltext Permission: restricted
Fulltext Availability: With Fulltext
Appears in Collections:SBS Student Reports (FYP/IA/PA/PI)

Files in This Item:
File Description SizeFormat 
KongKiatWhye.pdf
  Restricted Access
455.47 kBAdobe PDFView/Open

Page view(s) 50

324
Updated on Nov 28, 2020

Download(s)

4
Updated on Nov 28, 2020

Google ScholarTM

Check

Items in DR-NTU are protected by copyright, with all rights reserved, unless otherwise indicated.