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Isolation and Characterization of Genes That Affected the Growth of Burkholderia Species Mba4 by Transposon Mutagenesis

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Isolation and Characterization of Genes That Affected the Growth of Burkholderia Species Mba4 by Transposon Mutagenesis by Yun-Wing Faan
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This dissertation, "Isolation and Characterization of Genes That Affected the Growth of Burkholderia Species MBA4 by Transposon Mutagenesis" by 范殷榮, Yun-wing, Faan, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any way. We have altered the formatting in order to facilitate the ease of printing and reading of the dissertation. All rights not granted by the above license are retained by the author. Abstract: Abstract of thesis entitled ISOLATION AND CHARACTERIZATION OF GENES THAT AFFECTED THE GROWTH OF Burkholderia SPECIES MBA4 BY TRANSPOSON MUTAGENESIS Submitted by Faan Yun Wing for the degree of Master of Philosophy at The University of Hong Kong January 2008 This study focused on the isolation and characterization of genes that are essential for Burkholderia sp. MBA4 cells to grow on monochloroacetate. Three genes were identified by transposon mutagenesis to be necessary for the utilization of glycolate, which is the dehalogenation product of monochloroacetate. These genes encoded for a putative glycerate kinase, a putative LysR transcriptional regulator, and a putative glutathione synthetase. A mutant with its dehalogenase IVa gene (deh4a) being disrupted was also isolated. Genes that repressed the expression of deh4a in pyruvate-containing minimal medium were also identified by transposon Tn5-OT182-L. Tn5-OT182-L is a derivative of transposon Tn5-OT182 with the lacZ being removed. A plasmid pHKU1508 containing a lacZ was used to report the activity of the deh4a promoter. A gene encoding a putative two-component transcriptional regulator was found disrupted in mutant 15U09. This mutant showed ii 3-fold and 2-fold more deh4a transcripts in minimal medium containing only pyruvate and pyruvate plus monochloroacetate, respectively. A gene encoding a putative periplasmic component of an ABC-type iron transport system was observed disrupted in mutant 15U07. This mutant showed 6-fold and 2-fold more deh4a transcripts in minimal medium containing only pyruvate and pyruvate plus monochloroacetate, respectively. The growths of mutant 15U07 in the presence of 2, 2-dipyridyl and iron-limiting condition was better than the wild-type in iron-limiting medium. The sensitivities of the cells to streptonigrin implied that the mutant had increased intracellular iron content. iii DOI: 10.5353/th_b3955912 Subjects: Ralstonia solanacearumMutagenesisMolecular genetics
Release date NZ
January 27th, 2017
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Country of Publication
United States
Illustrations
colour illustrations
Imprint
Open Dissertation Press
Dimensions
216x279x8
ISBN-13
9781374677043
Product ID
26644189

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