Protocol 3
Methionine pathway inhibition
- Spin down the cells form 1 ml of
overnight culture in LB medium.
- Resuspend pellet in 1 ml of M9
medium [...] with carbon source (4 g/liter) and then add to 1 liter of
the some prewarmed medium.
- Cells grow up to mid-log phase.
- At mid-log phase add amino acids:
lysine, phenylalanine and threonine at 100 mg/liter; isoleucine and valine
at 50 mg/liter; and selenomethionine at 60 mg/liter.
- Induce the culture 15 minutes after
addition of the amino acids.
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Maciej Kozak's PPS'97 project
Protein engineering and its role in solving the phase
problem
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