Table 2.
Integration activities and DNA binding affinities of Int mutants
| Plasmid | Int variant | In vivo activity: % white/+ or – in subculture | In vitro activity: transformation assay (% white) | Int concentrations (nM) for 50% binding of probe |
|||
|---|---|---|---|---|---|---|---|
| attB | attP | attL | attR | ||||
| pHS62 | Int | 100/+ | 91 | 21–83 | 21–83 | 21 | 21 |
| pARM010 | hINT | 100/+ | 87 | 21–83 | 21–83 | 21 | 21 |
| pADD002 | hIntS374A | 42/+ | 78 | 83 | 21–83 | 5–21 | 5–21 |
| pARM024 | hIntS374L | 0/− | 2 | >1333 | 1333 | 1333 | 330 |
| pARM070 | hIntK375A | 2/+ | 20 | 333 | 83–333 | 83–333 | 83–333 |
| pARM103 | hIntK375Q | 0/− | <1 | 333 | 83 | 21–83 | 21 |
| pARM104 | hIntK375E | 0/− | <1 | 1333 | 83 | 83 | 21 |
| pARM071 | hIntR376A | 1/+ | <1 | 1333 | 83 | 83 | 21 |
| pARM105 | hIntR376E | 0/− | <1 | >1333 | >1333 | >1333 | >1333 |
| pARM096 | hIntR376P | 0/− | <1 | >1333 | 333 | 21 | 21 |
| pADD001 | hIntG377L | 61/+ | 78 | 21–83 | 21–83 | 21–83 | 21 |
| pADD010 | hIntG377A | 100/+ | 84 | 21–83 | 21–83 | 21–83 | 21–83 |
| pARM020 | hIntG377D | 40/+ | 58 | 21–83 | 21 | 5–21 | 5–21 |
| pARM021 | hIntG377F | 4/+ | <1 | 83–333 | 83–333 | 83–333 | 83 |
| pARM022 | hIntG377K | 0/− | 1 | 1333 | 21–83 | 83 | 21–83 |
| PARM072 | hIntE378A | 100/+ | 94 | 333 | 83–333 | 83–333 | 83–333 |
| pARM073 | hIntE379A | 100/+ | 93 | 83–333 | 83–333 | 83–333 | 83–333 |
| pARM074 | hIntS380A | 100/+ | 95 | 333 | 83–333 | 83–333 | 83–333 |
| pARM107 | hIntS380D | 32/+ | 29 | 21 | 21 | 5 | 5 |
| pARM108 | hIntS380N | 12/+ | 70 | 5–21 | 5 | 5 | 1.3 |
| pARM109 | hIntS380T | 100/+ | 94 | 1.3–5 | 5 | 1.3–5 | 1.3 |
| pARM100 | hIntI381M | 100/+ | 95 | ND | ND | ND | ND |
| pARM106 | hIntV371SUGA | 0/− | <1 | >1333 | 333–1331 | >1333 | >1333 |
Bold indicates a lower affinity for a substrate than the wild-type Int. ‘ND’ indicates that the data were not determined.