Table 1.
Oligonucleotides that were used for site specific mutagenesis. Lowercase indicates altered bases. Only the forward primer is shown.
| Product Name | Start Plasmid | Synthetic Oligonucleotide | Description | Calculated Molecular Wt |
|---|---|---|---|---|
| Wild type | A (Wt NHE HA tag) | N/A | 815-HA | 95220 |
| Wild type | B (WT NHE1 GFP tag) | N/A | 815-GFP | 121612 |
| NHE-700 | A | 5′-CCATGTCTCGGGCCCtCgagGGCTCAGACCCACTG-3′ | Stop @AA 700-HA | 81187 |
| NHE-675 | A | 5′-GGCAGAAGGCCCGGCtcgaGGAGCAGAAGATCAAC-3′ | Stop @AA 675-HA | 80229 |
| NHE-625 | A | 5′-CTGCCAGCACTGTCCctcGAgAAGGAGGAGGAGATC-3′ | Stop @AA 625-HA | 73953 |
| NHE-600 | A | 5′-GGCAAGATCCCCTCTctCGagTCCACCGTCTCCATG CAG-3′ | Stop @AA 600-HA | 71295 |
| NHE-582 | A | 5′-CCACAAGATGGAGATGctcgAGGCCATCGAGCTGGTG-3′ | Stop @AA 582-HA | 69569 |
| NHE-566 | A | 5′-GTGTCTGATAGCTGGCGAGCtCgagAAGGAGCCCCAGC TCATTG-3′ | Stop @AA 566-HA | 67579 |
| NHE-564 | A | 5′-GTGAAGAAGTGTCTGATAGCTctcGAGCGCTCCAAG GAGCCCCAGCTC-3′ | Stop @AA 564-HA | 67393 |
| NHE-562 | A | 5′-GAAATATGTGAAGAAGTGTCTcgagGCTGGCGAGC GCTCCAAG-3′ | Stop @AA 562-HA | 67096 |
| NHE-E566A | A | 5′-GTGTCTGATAGCTGGCgcGctggAGgatgacGGCCGCATCT TTTACCC-3′ | NHE-566 with last AA as A-HA | 67521 |
| NHE-E566K | A | 5′-GTGTCTGATAGCTGGCaAGctcgAGgatgacGGCCGCATC TTTTACCC-3′ | NHE-566 with last 4 AA as K-HA | 67578 |
| NHE-563-6A | A | 5′-ATATGTGAAGAAGTGTCTGgcAGCTgcCgcGctggAGgatgacGGCCGCATCTTTTACC-3′ | NHE-566 with last 4 AA as A-HA | 67493 |
| NHE-562-GFP | B | 5′-GAAATATGTGAAGAAGTGTCTcgagGCTGGCGAGC GCTCCAAG-3′ | Stop @AA 562-GFP | 90038 |
| NHE-564-GFP | B | 5′-GTGAAGAAGTGTCTGATAGCTctcGAGCGCTCCAAG GAGCCCCAGCTC-3′ | Stop @AA 564-GFP | 90335 |
| NHE-566-GFP | B | 5′-GTGTCTGATAGCTGGCGAGCtCgagAAGGAGCCCCAGCTCATTG-3′ | Stop @AA 566-GFP | 90521 |
A, alanine; AA, amino acids; GFP-green fluorescent protein tag; HA-hemagglutinin tag.