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Pribnow box (-10) T 89 A 99 T 50 A 65 A 65 T 100  70 binding site (-35) T 85 T 83 G 81 A 61 C 69 A 52.

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Presentation on theme: "Pribnow box (-10) T 89 A 99 T 50 A 65 A 65 T 100  70 binding site (-35) T 85 T 83 G 81 A 61 C 69 A 52."— Presentation transcript:

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3 Pribnow box (-10) T 89 A 99 T 50 A 65 A 65 T 100  70 binding site (-35) T 85 T 83 G 81 A 61 C 69 A 52

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18 pMAL vectors : ColE1 ori ; M13 ori ; Ptac ; rrnB terminators ; lacI q ; bla Fusion construct : malE - DDDDDDDDDD - IEGR - MCS - lacZ  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 gt11 (lacZ) factor Xa I - E - G - R * factor Xa I - E - G - R * XmnI nnn nnn nGA Ann nnT TCn nnn XmnI nnn nnn nGA Ann nnT TCn nnn ATC-GAG-GGA-AGG-ATT-TCA-GAA-TTC- ATC-GAG-GGA-AGG-ATT-TCA-GAA-TTC- EcoRI EcoRI

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