Part:BBa_J100198:Experience
This part works as designed. thyA- cells that contain J100198 do not grow unless theophylline is added to the media. Growth rates are comparable with 0.3, 0.6 and 1.25 mM theophylline added to LB broth + 50 µg/mL ampicillin (to select for pSB1A8 plasmid) + 50 µg/mL kanamycin (to select for thyA- mutation). J100198 is an improvement over part Part:BBa_J100136 which failed because it permitted growth in the absence of theophylline.
Photograph of thyA- cells that contain J100198 in pSB1A8 grown in LB broth + 50 µg/mL ampicillin (to select for pSB1A8 plasmid) + 50 µg/mL kanamycin (to select for thyA- mutation). From left to right: 0, 0.3, 0.6 and 1.25 mM theophylline.
OD590 of thyA- cells that contain J100198 in pSB1A8 grown in LB broth + 50 µg/mL ampicillin (to select for pSB1A8 plasmid) + 50 µg/mL kanamycin (to select for thyA- mutation) with added theophylline as indicated. The cells were grown overnight and measured then ext morning.
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