Table 1 Oligos applied in this study. I: Synthesized DNA templates bearing either wild-type binding site (Zif_1) for zif268 or one of its variants (Zif_2 to Zif_6) used for generating DNA binding sites by PCR amplification, where KS-1 and SK-1 are two primers (low case). II: Oligos employed to construct five zif268 variants with QuickChangeâ„¢ XL site-directed mutagenesis Kit (Stratagene) using pzif268 as a template.
I | Zif_1 tcgaggtcgacggtatcGCGTGGGCGCtccactagttctagagcggccgccac |
|---|---|
| Â | Zif_2 tcgaggtcgacggtatcGCGTGGGCACtccactagttctagagcggccgccac |
| Â | Zif_3 tcgaggtcgacggtatcGCGTGGGCCCtccactagttctagagcggccgccac |
| Â | Zif_4 tcgaggtcgacggtatcGCGTGGGAGCtccactagttctagagcggccgccac |
| Â | Zif_5 tcgaggtcgacggtatcGCGTGGGAACtccactagttctagagcggccgccac |
| Â | Zif_6 tcgaggtcgacggtatcGCGTGGGACCtccactagttctagagcggccgccac |
| Â | KS-1 tcgaggtcgacggtatc |
| Â | SK*-1 gtggcggccgctctagaact (SK-1 was fluorescent labeled with either FAM, HEX, TAMRA, ROX, or CY5) |
II | 18Q_plus 5' CGCCGCTTTTCTcagTCGGATGAGCTTACCCGCC |
| Â | 18Q_minus 5' GGCGGGTAAGCTCATCCGActgAGAAAAGCGGCG |
| Â | 18D_plus 5' CGCCGCTTTTCTgatTCGGATGAGCTTACCCGCC |
| Â | 18D_minus 5' GGCGGGTAAGCTCATCCGAatcAGAAAAGCGGCG |
| Â | 21N_plus 5' CGCCGCTTTTCTCGCTCGGATaacCTTACCCGCC |
| Â | 21N_minus 5' GGCGGGTAAGgttATCCGAGCGAGAAAAGCGGCG |
| Â | 18Q_21N_plus 5' CGCCGCTTTTCTcagTCGGATaacCTTACCCGCC |
| Â | 18Q_21N_minus 5' GGCGGGTAAGgttATCCGActgAGAAAAGCGGCG |
| Â | 18D_21N_plus 5' CGCCGCTTTTCTgatTCGGATaacCTTACCCGCC |
| Â | 18D_21N_minus 5' GGCGGGTAAGgttATCCGAatcAGAAAAGCGGCG |