Protocol 2


Cell growth in selenometionine


  1. Isolate single colonies by streaking an LB plates supplemented with antibiotics with strain of interest. Incubate at 37 degree.
  2. Ferment the cells in media containing all appropriate antibiotics
  1. Induce specific cloned protein synthesis.
  2. Harvest the cells in mid- to late-log phase. Resuspend the harvested cells in appropriate lysis buffer and freeze with dry ice. Store at -80 degrees.
Ingredients Concentration
1. Minimal medium

2. All amino acids except methionine

3. Adenine, guanine, thymine and uracil

4. Selenomethionine

5. LB

Minimal medium A [27], with carbon source at 5g/l

40 mg/l

0.5 g/l

10-50 mg/l

for 100 ml starter 5% (v/v) LB for 20 l fermenter

      Ingredients Concentration
      1. Minimal medium

      2. All amino acids except methionine

      3. Adenine, guanine, thymine and uracil

      4. Selenomethionine

      5. Thiamine

      6. Biotin (optional)

      Minimal medium A [27], with carbon source at 5g/l

      40 mg/l

      0.5 g/l

      10-50 mg/l

      2 mg/l

      2 mg/l


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Maciej Kozak's PPS'97 project
Protein engineering and its role in solving the phase problem
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