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

Pribnow box (-10)

T89 A99 T50 A65 A65 T100

70 binding site (-35)

T85 T83 G81 A61 C69 A52

pMAL vectors :

ColE1 ori ; M13 ori ; Ptac ; rrnB terminators ; lacIq ; 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 fusion in XmnXmnI : exact joining of target protein at factor Xa sequenceI : exact joining of target protein at factor Xa sequence 10 Asp residues (D) separate the two fusion moieties10 Asp residues (D) separate the two fusion moieties insertion in insertion in EcoEcoRI is identical (in reading frame) as in RI is identical (in reading frame) as in gt11 (gt11 (lacZlacZ))

factor Xa I - E - G - R *factor Xa I - E - G - R *

XmnXmnI nnn nnn nI nnn nnn nGA AGA Ann nnnn nnT TCT TCn nnn n nnn ATC-GAG-GATC-GAG-GGA-AGA-AGG-ATGG-ATT-TCT-TCA-A-GAA-TTCGAA-TTC-- EcoEcoRI RI