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DNA shuffling of biphenyl dioxygenase genes, bphAE or bphE, from Comamonas testosteroni B-356 and Burkholderia cepacia LB400

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DNA shuffling of biphenyl dioxygenase genes, bphAE or bphE, from Comamonas testosteroni B-356 and Burkholderia cepacia LB400

Aumont, Roch (2002) DNA shuffling of biphenyl dioxygenase genes, bphAE or bphE, from Comamonas testosteroni B-356 and Burkholderia cepacia LB400. Masters thesis, Concordia University.

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Abstract

Bacterial degradation of biphenyl and some polychlorinated biphenyls is initiated by and dependent on the ability of biphenyl 2,3-dioxygenase to dihydroxylate substrate. Biphenyl 2,3-dioxygenase is a multi-component enzyme requiring reductase (BphG) and ferredoxin (BphF) components that transport electrons from NADH to the terminal dioxygenase. DNA shuffling of bphAE , using various techniques to create random fragments, resulted either in a very low yield of reassembled product and a high level of non-specific recombination, or in reassembly of wild type enzymes. Sequencing of parts of 13 randomly chosen active (8) or inactive (5) shuffled bphAE genes revealed only two recombination events. Inability to shuffle both subunits Ì and Ý together may be caused by the highly variable inter-genic region, which may not allow homologous recombination to occur between DNA templates. A second strategy was DNA shuffling of the Ý-subunit alone, as this subunit has been shown to affect enzyme specificity

Divisions:Concordia University > Faculty of Arts and Science > Chemistry and Biochemistry
Item Type:Thesis (Masters)
Authors:Aumont, Roch
Pagination:xii, 93 leaves : ill. (some col.) ; 29 cm.
Institution:Concordia University
Degree Name:M.Sc.
Program:Chemistry
Date:2002
Thesis Supervisor(s):Powlowski, Justin B
Identification Number:TD 192.5 A86 2002
ID Code:2213
Deposited By: Concordia University Library
Deposited On:27 Aug 2009 17:26
Last Modified:13 Jul 2020 19:51
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