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Molecular studies of the sdaA and sdaB genes and their gene products in Escherichia coli K-12

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Molecular studies of the sdaA and sdaB genes and their gene products in Escherichia coli K-12

Su, Hongsheng (1991) Molecular studies of the sdaA and sdaB genes and their gene products in Escherichia coli K-12. PhD thesis, Concordia University.

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Abstract

This work demonstrates the existence of Escherichia coli K-12 of two L-serine deaminating enzymes, L-serine deaminase (L-SD)#1 and L-SD#2. It demonstrates that the structure of L-SD#1 is coded by the sdaA gene, which has been cloned and sequenced. The entire sdaA gene was fused to the lacZ gene by mutating the stop-codon of sdaA and ligating in-frame to lacZ. The fused gene directed the formation of a large protein showing both L-SD and $\beta$-galactosidase activities. L-SD#1 has been extensively purified for the first time by use of a three-part fusion protein and some of its characteristics studied. L-SD#2 is synthesized in wild-type cells in LB medium. A mutation in the sdaX gene established its expression in minimal medium. An insertion in sdaB abolished L-SD#2 activity in an sdaA::Cm$\sp{\rm r}$ sdaX strain, allowing the sdaB gene to be cloned by restoring growth on L-serine. The sdaA gene was located at 41 minutes; sdaB and sdaX both were located near 60.1 minutes and may be the same gene. Some experiments directed towards the identification of the metabolic role of L-SD are included.

Divisions:Concordia University > School of Graduate Studies
Item Type:Thesis (PhD)
Authors:Su, Hongsheng
Pagination:xvi, 212 leaves : ill. (some mounted) ; 29 cm.
Institution:Concordia University
Degree Name:Ph. D.
Program:Special Individual Program
Date:1991
Thesis Supervisor(s):Newman, E. B
Identification Number:QR 82 E6S8 1991
ID Code:26
Deposited By: Concordia University Library
Deposited On:27 Aug 2009 17:09
Last Modified:13 Jul 2020 19:45
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