TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristicsa | Source or reference |
|---|---|---|
| E. coli strains | ||
| BL21 (DE3)(pLysS) | hsdS gal (λcIts857 ind1 Sam7 nin5 lacUV5-T7 gene 1) pLysS (Cmr) | 38, 54 |
| DH5α | F− φ80dlacZΔM15 Δ(lacZYA-argF)U169 endA1 recA1 hsdR17 (rK− mK+) deoR thi-1 supE44 λ− gyrA96 relA1 | GIBCO BRL |
| TG1 | supE hsdΔ5 thi Δ(lac-proAB) F′ (traD36 proAB+ lacIq lacZΔM15) | Amersham Sculptor-Kit (17, 38) |
| Plasmids | ||
| pPX31 | pUC19 (Apr) with catB from P. putida PRS2000 on a 2.1-kbp PstI fragment | L. N. Ornston (24) |
| pET-11a* | Apr PT7lac derivative of pET-11a containing additional KpnI, XhoI, and SalI cloning sites | J. Altenbuchner (54) |
| pUC BM20 | Apr Plac derivative of pUC18 with larger multiple cloning site | Boehringer Mannheim |
| pBluescript II KS(+) | Apr Plac f1(+) and ColE1 origin | Stratagene (2) |
| pBluescript II SK(+) | Apr Plac f1(+) and ColE1 origin | Stratagene (2) |
| pCATB1 | pET-11a* (Apr) with catB flanked by NdeI and BamHI sites | This study |
| pCATB2 | pUC BM20 (Apr) with catB flanked by NdeI and BamHI sites | This study |
| pCATB3 | pUC BM20 (Apr) with 855-bp HindIII-BamHI fragment of catB | This study |
| pCATB4 | pBluescript II KS(+) (Apr) with 237-bp SalI-HindIII fragment of catB | This study |
| pCATB5 | pBluescript II SK(+) (Apr) with 211-bp SacII-XhoI fragment of catB | This study |
| pCATB6 | pBluescript II SK(+) (Apr) with 319-bp SacII-HincII fragment of catB | This study |
| pCATB51 | pET-11a* (Apr) with NdeI-BamHI fragment carrying I54V-CatB | This study |
| pCATB52 | pET-11a* (Apr) with NdeI-BamHI fragment carrying Y59W-CatB | This study |
| pCATB53 | pET-11a* (Apr) with NdeI-BamHI fragment carrying K276N-CatB | This study |
| pCATB54 | pET-11a* (Apr) with NdeI-BamHI fragment carrying A271S-CatB | This study |
| pCATB55 | pET-11a* (Apr) with NdeI-BamHI fragment carrying F329I-CatB | This study |
| pCATB56 | pET-11a* (Apr) with NdeI-BamHI fragment carrying L333V-CatB | This study |
| pCATB57 | pET-11a* (Apr) with NdeI-BamHI fragment carrying I54V-Y59W-CatB | This study |
| pCATB58 | pET-11a* (Apr) with NdeI-BamHI fragment carrying I54V-F329I-CatB | This study |
| pCATB59 | pET-11a* (Apr) with NdeI-BamHI fragment carrying Y59-F329I-CatB | This study |
| pCATB60 | pET-11a* (Apr) with NdeI-BamHI fragment carrying I54V-Y59W-F329I-CatB | This study |
a
Apr, resistance to ampicillin; Cmr, resistance to chloramphenicol.