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...
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| Other Authors: | |
| Format: | Final Year Project (FYP) |
| Language: | English |
| Published: |
2009
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| Online Access: | http://hdl.handle.net/10356/16341 |
| _version_ | 1826115041770864640 |
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| author | Kong, Kiat Whye. |
| author2 | Sze Chun Chau |
| author_facet | Sze Chun Chau Kong, Kiat Whye. |
| author_sort | Kong, Kiat Whye. |
| collection | NTU |
| 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. |
| first_indexed | 2024-10-01T03:49:04Z |
| format | Final Year Project (FYP) |
| id | ntu-10356/16341 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T03:49:04Z |
| publishDate | 2009 |
| record_format | dspace |
| spelling | ntu-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 |
| 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. |
| 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_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_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_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 |
| topic | DRNTU::Science::Biological sciences::Microbiology::Bacteria |
| url | http://hdl.handle.net/10356/16341 |
| work_keys_str_mv | AT kongkiatwhye optimizationofconditionsrequiredforthecharacterizationofescherichiacolipromotersresponsivetointerspeciesinteractionwithcommensalpartnersklebsiellapneumoniaeandenterococcusfaecalis |