TABLE 1.
Bacterial strains and plasmids used in this study
| Strain/plasmid | Genotype/phenotype/descriptionb | Reference or source |
|---|---|---|
| A. hydrophila strains | ||
| Ah65 | Wild type | 14 |
| C5.84 | Ah65 exeA::Tn5-751; Kmr | 13 |
| Plasmids | ||
| pMMB207 | tac promoter, wide-host-range vector; Cmr | 22 |
| pKRJ50.2 | 2.5-kb exeAB BstXI in SmaI of pBluescript II KS(+); Ampr | 13 |
| pKRJ50.2RE | pKRJ50.2 with silent mutations for RsrII, AfilII and BspEIa | This study |
| pRJ31.1 | 2.5-kb exeAB BstXI in SmaI of pMMB207; Cmr | 13 |
| pRJ31.1 F476A | PRJ31.1 encoding ExeA F476A | This study |
| pRJ31.1 K481A | pRJ31.1 encoding ExeA F487A | This study |
| pRJ31.1I F487A | pRJ31.1 encoding ExeA F487A | This study |
| pRJ31.1Q488A | pRJ31.1 encoding ExeA Q488A | This study |
| pRJ31.1D496A | pRJ31.1 encoding ExeA D496A | This study |
| pRJ31.1G497A | pRJ31.1 encoding ExeA G497A | This study |
| pRJ31.1I498W | pRJ31.1 encoding ExeA I498W | This study |
| pRJ31.1T503A | pRJ31.1 encoding ExeA T503A | This study |
| pRJ31.1L507A | pRJ31.1 encoding ExeA L507A | This study |
| pET30a | Expression vector; T7lac promoter; Kmr | Novagen |
| pP-ExeA | N-His P-ExeA fragment in NdeI/XhoI of pET30a | 18 |
| pP-ExeAF476A | pP-ExeA encoding P-ExeAF476A | This study |
| pP-ExeAK481A | pP-ExeA encoding P-ExeAK487A | This study |
| pP-ExeAF487A | pP-ExeA encoding P-ExeAF487A | This study |
| pP-ExeAD496A | pP-ExeA encoding P-ExeAD496A | This study |
a
See Table 2 for locations of the three restriction sites.
b
Cmr, chloramphenicol resistance; Ampr, ampicillin resistance; Kmr, kanamycin resistance.