Construct | Primers | Restriction endonucleases | Protein product |
---|---|---|---|
pAPA' | N-terminal: 5' CCA GAA TTC CAT ATGÂ ACC GTC GAC CAG GCG GCG GAG C 3' C-terminal: 5' CCA GGA TCC TTA GTT GGC CCC GTA GAG CTT CTG G 3' | Nde I Bam HI | AprA catalytic fragment (APA') |
pPelAPA' | N-terminal: 5' CCG AAT TCC ATG GÂ CC GTC GCC GTC GAC CAG GCG GCG GAG C 3' C-terminal: Same as pAPA' | Nco I | APA' with pelB secretion signal |
pNusAPA' | N-terminal: 5' G GGC GGG GAC GAA TTG GTC AAT GGC 3' C-terminal: Same as pAPA' | Blunt end ligation at N-terminus: no digestion required | APA' with NusA solubility tag and His6 tag |
pWApr | N-termnal: 5' CCA GAA TTC CAT ATGÂ GGT CGT AGC GAT GCG TAT ACC 3' C-terminal: 5' CCA GGA TCCÂ TCA GAC GAC GAT GTC GGC CTG G 3' | Nde I Bam HI | Mature AprA without pro-peptide |