Presentation Title

Transposon Mutagenesis of Transporter Gene Affects Motility and EPS in Paraburkholderia unamae

Faculty Mentor

Dr. Shelley Thai

Start Date

17-11-2018 12:30 PM

End Date

17-11-2018 2:30 PM

Location

HARBESON 10

Session

POSTER 2

Type of Presentation

Poster

Subject Area

biological_agricultural_sciences

Abstract

Intro: The model organism of this study, Paraburkholderia unamae is a gram-negative, highly motile bacterium that is non-pathogenic, diazotrophic and can be found in maize, sugarcane, and coffee. This research was conducted to study the genes of P. unamae responsible for the motility and production of exopolysaccharides (EPS). The latter helps bacteria to adhere to the surfaces of plants and protects them from desiccation. The motility of P. unamae contributes to the maximization of its survival and its ability to search for a favorable environment that contains nutrients. Objective: The objective of this experiment was to study the genes that contribute to the reduction of motility and overexpression of EPS and to understand the positional effect of the transposon insertion on the targeted gene expression. Methods: To randomly disrupt genes, transposon mutagenesis was performed by introducing the vector pRL27 that contains the transposable element into P. unamae. Of 1000 screened for overexpression of EPS and reduced motility, 5 mutants were isolated to perform gene analysis. Results: Genetic analysis determined that the transposon was inserted in the intergenic region of the cation diffusion facilitator family transporter and H-NS histone family gene. The results of this study indicate that motility decreases as the transposon nears the cation diffusion facilitator family transporter gene. There is also a negative correlation between the mutant motility and the amount of EPS produced. Conclusion: The reason for the gradual decrease in motility in this study is hypothesized to be the result of the presence of enhancer-like and silencer-like regions of cation diffusion facilitator family transporter gene that could be located in the intergenic region. The interruption of these two regions alters the function of the gene, which in turn affects the phenotypic characteristics.

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Nov 17th, 12:30 PM Nov 17th, 2:30 PM

Transposon Mutagenesis of Transporter Gene Affects Motility and EPS in Paraburkholderia unamae

HARBESON 10

Intro: The model organism of this study, Paraburkholderia unamae is a gram-negative, highly motile bacterium that is non-pathogenic, diazotrophic and can be found in maize, sugarcane, and coffee. This research was conducted to study the genes of P. unamae responsible for the motility and production of exopolysaccharides (EPS). The latter helps bacteria to adhere to the surfaces of plants and protects them from desiccation. The motility of P. unamae contributes to the maximization of its survival and its ability to search for a favorable environment that contains nutrients. Objective: The objective of this experiment was to study the genes that contribute to the reduction of motility and overexpression of EPS and to understand the positional effect of the transposon insertion on the targeted gene expression. Methods: To randomly disrupt genes, transposon mutagenesis was performed by introducing the vector pRL27 that contains the transposable element into P. unamae. Of 1000 screened for overexpression of EPS and reduced motility, 5 mutants were isolated to perform gene analysis. Results: Genetic analysis determined that the transposon was inserted in the intergenic region of the cation diffusion facilitator family transporter and H-NS histone family gene. The results of this study indicate that motility decreases as the transposon nears the cation diffusion facilitator family transporter gene. There is also a negative correlation between the mutant motility and the amount of EPS produced. Conclusion: The reason for the gradual decrease in motility in this study is hypothesized to be the result of the presence of enhancer-like and silencer-like regions of cation diffusion facilitator family transporter gene that could be located in the intergenic region. The interruption of these two regions alters the function of the gene, which in turn affects the phenotypic characteristics.