Presentation Title

Disruption of the Genes fucP and rpoD Caused Overexpression of EPS in Paraburkholderia unamae

Faculty Mentor

Shelley Thai Ph.D., Michelle Lum Ph.D.

Start Date

17-11-2018 8:30 AM

End Date

17-11-2018 10:30 AM

Location

CREVELING 34

Session

POSTER 1

Type of Presentation

Poster

Subject Area

biological_agricultural_sciences

Abstract

Disruption of the Genes fucP and rpoD Caused Overexpression of EPS in Paraburkholderia unamae

Author: Meghri Shaverdian, Glendale Community College; Melanis Ghadimi, Glendale Community College

Mentor: Dr. Shelley Thai, Biology Division, Glendale Community College; Dr. Michelle Lum, Department of Biology, Loyola Marymount University

Paraburkholderia unamae is a non-pathogenic plant-associative bacterium. These bacteria can be used for nitrogen fixation, replacing fertilizers and other chemicals which may harm the environment. The main objective is to study the genes involved in exopolysaccharide (EPS) production and motility by generating random mutations throughout the genome using Tn5 transposon in the process of transposon mutagenesis. After conjugation, mutants were screened for alternation in EPS and motility phenotypes. Plasmids carrying the transposon were isolated and sequenced. Both mutants MS 4-42 and MelG 1-39 exhibited an over-EPS production with a moderate increase in motility. According to DNA sequencing data for mutants MS 4-42 and MelG 1-39, the Tn5 transposon insertion disrupted the genes fucP and rpoD, which encode for fucose permease and RNA polymerase sigma 70 subunit, respectively. Fucose permease is a transmembrane protein involved in regulating the concentration of fucose. We hypothesize that disruption of fucP resulted in the inability of fucose to be exported. This over-accumulation of fucose in a cell can lead to over-production of EPS. RNA polymerase σ70 subunit binds to RNA polymerase to initiate transcription. Alternative sigma factors, such as RNA polymerase sigma-H factor, AlgU, exist to start transcription when major sigma factor σ70 is not available. AlgU is responsible for activating the production of alginate, a polysaccharide component in the EPS of some bacteria. We hypothesize that disruption of σ70 affected the production of alginate and the overproduction of EPS. Further studies are required to further understand the effect of the disrupted genes in the production of EPS in P. unamae.

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Nov 17th, 8:30 AM Nov 17th, 10:30 AM

Disruption of the Genes fucP and rpoD Caused Overexpression of EPS in Paraburkholderia unamae

CREVELING 34

Disruption of the Genes fucP and rpoD Caused Overexpression of EPS in Paraburkholderia unamae

Author: Meghri Shaverdian, Glendale Community College; Melanis Ghadimi, Glendale Community College

Mentor: Dr. Shelley Thai, Biology Division, Glendale Community College; Dr. Michelle Lum, Department of Biology, Loyola Marymount University

Paraburkholderia unamae is a non-pathogenic plant-associative bacterium. These bacteria can be used for nitrogen fixation, replacing fertilizers and other chemicals which may harm the environment. The main objective is to study the genes involved in exopolysaccharide (EPS) production and motility by generating random mutations throughout the genome using Tn5 transposon in the process of transposon mutagenesis. After conjugation, mutants were screened for alternation in EPS and motility phenotypes. Plasmids carrying the transposon were isolated and sequenced. Both mutants MS 4-42 and MelG 1-39 exhibited an over-EPS production with a moderate increase in motility. According to DNA sequencing data for mutants MS 4-42 and MelG 1-39, the Tn5 transposon insertion disrupted the genes fucP and rpoD, which encode for fucose permease and RNA polymerase sigma 70 subunit, respectively. Fucose permease is a transmembrane protein involved in regulating the concentration of fucose. We hypothesize that disruption of fucP resulted in the inability of fucose to be exported. This over-accumulation of fucose in a cell can lead to over-production of EPS. RNA polymerase σ70 subunit binds to RNA polymerase to initiate transcription. Alternative sigma factors, such as RNA polymerase sigma-H factor, AlgU, exist to start transcription when major sigma factor σ70 is not available. AlgU is responsible for activating the production of alginate, a polysaccharide component in the EPS of some bacteria. We hypothesize that disruption of σ70 affected the production of alginate and the overproduction of EPS. Further studies are required to further understand the effect of the disrupted genes in the production of EPS in P. unamae.