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Jang Ki-Hyo,Britz Margaret L. The Korean Society for Biotechnology and Bioengine 2005 Biotechnology and Bioprocess Engineering Vol.10 No.3
To investigate impediments to plasmid transformation in Brevibacterium flavum BF4 and B. lactofermentum BL1, cell surface barriers were determined by measuring growth inhibition whilst enzymatic barriers were determined by comparing DNA methylation properties. B. lactofermentum was more sensitive to growth inhibition by glycine than B. flavum. Release of cellular proteins during sonication was more rapid for B. lactofermentum than for B. flavum. Plasmid DNA (pCSL 17) isolated from B. flavum transformed recipient $McrBC^+$ strains of Escherichia coli with lower efficiency than $McrBC^-$. McrBC digestion of this DNA confirmed that B. flavum contain methylated cytidines in the target sequence of McrBc sequences but B. lactofermentum contained a different methylation pattern. DNA derived from the B. lactofermentum transformed recipient $EcoKR^+$ strains of E. coli with lower efficiency than $EcoKR^-$, indicating the presence of methylated adenosines in the target sequence of EcoK sequences. The present data describe the differences in the physical and enzymatic barriers between two species of corynebacteria and also provide some insight into the successful foreign gene expression in corynebacteria.
Ki-Hyo Jang,Margaret L. Britz 한국생물공학회 2005 Biotechnology and Bioprocess Engineering Vol.10 No.3
To investigate impediments to plasmid transformation in Brevibacterium flavum BF4 and B. lactofermentum BL1, cell surface barriers were determined by measuring growth inhibition whilst enzymatic barriers were determined by comparing DNA methylation properties. B. lactofermentum was more sensitive to growth inhibition by glycine than B. flavum. Release of cellular proteins during sonication was more rapid for B. lactofermentum than for B. flavum. Plasmid DNA (pCSL17) isolated from B. flavum transformed recipient McrBC+ strains of Escherichia coli with lower efficiency than McrBC. McrBC digestion of this DNA confirmed that B. flavum contain methylated cytidines in the target sequence of McrBC sequences but B. lactofermentum contained a different methylation pattern. DNA derived from the B. lactofermentum transformed recipient EcoKR+ strains of E. coli with lower efficiency than EcoKR, indicating the presence of methylated adenosines in the target sequence of EcoK sequences. The present data describe the differences in the physical and enzymatic barriers between two species of corynebacteria and also provide some insight into the successful foreign gene expression in corynebacteria.
Marzieh Hosseini Nezhad,Matthew Knight,Margaret Lorraine Britz 한국식품과학회 2012 Food Science and Biotechnology Vol.21 No.1
In this research, changes were observed in cell surface proteins of a typical strain of the Lactobacillus casei was investigated in response to acidic growth conditions. Two dimensional electrophoresis and Western blot analyses were carried out to detect changes in relative abundance of proteins at the cell surface. The identity of the differentially expressed proteins extracted by LiCl, a chemical routinely used to extract surface proteins of lactic acid bacteria, was determined by MALDI-TOF/TOF MS. Many enzymes involved in glycolysis were up-regulated in the cell surface fraction following growth at low pH, including enolase,lactate dehydrogenase, and glyceraldehyde-3-phosphate dehydrogenase. Several of these proteins were also related to adhesion and generalized stress responses. It is demonstrated that growth of L. casei under acidic conditions caused molecular changes at the cell surface to develop an adaptive strategy corresponding to slower growth at low pH.