Difference between revisions of "Part:BBa K220000:Experience"
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Since DMG alone did not rescue growth of control cells it was hypothesized that cells supplemented with DMG had basal levels of enzyme due to a leaky promoter. These cells may have grown best because they were able to convert DMG into gylcine betaine, but were not over producing the protein. Work was begun on attenuating expression by using a lower copy number plasmid with a weaker promoter. | Since DMG alone did not rescue growth of control cells it was hypothesized that cells supplemented with DMG had basal levels of enzyme due to a leaky promoter. These cells may have grown best because they were able to convert DMG into gylcine betaine, but were not over producing the protein. Work was begun on attenuating expression by using a lower copy number plasmid with a weaker promoter. | ||
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===Applications of BBa_K220000=== | ===Applications of BBa_K220000=== | ||
Latest revision as of 14:16, 2 November 2009
!!! WARNING: Pst1 cut site within gene !!!
http://i33.tinypic.com/sbmrz7.png
This graph shows the growth of DH10B control cells in .6 M NaCl/LB. Cells were treated with 1mM IPTG, 1 mM dimethyl gylcine (DMG), both, or nothing. These treatments had little effect on control cells.
http://i34.tinypic.com/4qera.png
This graph shows the growth of DH10B cells harboring the GsSDMT gene on an expression plasmid in .6 M NaCl/LB. Cells were treated with 1mM IPTG, 1 mM dimethyl gylcine (DMG), both, or nothing. Induction caused a decrease of growth due to the high strength of the promoter and associated stress of production. The expression of the gene helped growth when DMG was supplied relative to IPTG only. Since DMG alone did not rescue growth of control cells it was hypothesized that cells supplemented with DMG had basal levels of enzyme due to a leaky promoter. These cells may have grown best because they were able to convert DMG into gylcine betaine, but were not over producing the protein. Work was begun on attenuating expression by using a lower copy number plasmid with a weaker promoter.
Applications of BBa_K220000
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