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Pribnow box (-10) T 89 A 99 T 50 A 65 A 65 T 100 s 70 binding site (-35)

Pribnow box (-10) T 89 A 99 T 50 A 65 A 65 T 100 s 70 binding site (-35) T 85 T 83 G 81 A 61 C 69 A 52. pMAL vectors : ColE1 ori ; M13 ori ; Ptac ; rrnB terminators ; lacI q ; bla Fusion construct : malE - DDDDDDDDDD - IEGR - MCS - lacZ a

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Pribnow box (-10) T 89 A 99 T 50 A 65 A 65 T 100 s 70 binding site (-35)

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  1. Pribnow box (-10) T89 A99 T50 A65 A65 T100 s70 binding site (-35) T85 T83 G81 A61 C69 A52

  2. pMAL vectors : • ColE1 ori ; M13 ori ; Ptac ; rrnB terminators ; lacIq ; bla • Fusion construct : malE - DDDDDDDDDD - IEGR - MCS - lacZa • pMAL-c2 : malE signal peptide sequence deleted • pMAL-p2 : malE signal peptide sequence : secretion to periplasm • fusion in XmnI : exact joining of target protein at factor Xa sequence • 10 Asp residues (D) separate the two fusion moieties • insertion in EcoRI is identical (in reading frame) as in lgt11 (lacZ) factor Xa I - E - G - R * XmnI nnn nnn nGA Ann nnT TCn nnn ATC-GAG-GGA-AGG-ATT-TCA-GAA-TTC- EcoRI

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