Table 2. Bacterial strains and plasmids used in this study.
Strain or plasmid | Relevant characteristic(s) | Source |
Bacillus licheniformis 749/I | β-lactamase inducible strain, wild-type | |
Bacillus subtilis 168 | Recipient cell for E. coli/Bacillus shuttle vectors | ATCC 23857 |
Escherichia coli GI724 | Host cell for heterologous expression of BlaR1 L3, F-, lamda-, lacPL8, ampC::Ptrpcl | Invitrogen |
pUCBM20 | Cloning plasmid, Apr | Boehringer |
pCR-ScriptTMSK(+) | Cloning vector for PCR fragments, Apr | Stratagene |
pLEX | Expression vector from inducible P-lambda promoter, Apr | Novagen |
pDML995 | A derivative of the Bacillus/E. coli shuttle vector pMK4 [39] carrying the wild-type B. licheniformis 749I bladivergon, Apr in E.coli and Cmr in Bacillus | A. Brans (unpublished) |
pDML1251 | pUCBM20 derivative carrying the SstI/EcoRIblaR1 fragment | This study |
pDML1255 | pDML1251 derivative carrying BlaR1H212A/E213A/H216A mutation | This study |
pDML1256 | pDML1251 derivative carrying BlaR1 E213A mutation | This study |
pDML1257 | pDML1251 derivative carrying BlaR1 H212A/H216A mutation | This study |
pDML1258 | pDML1251 derivative carrying BlaR1 H216A mutation | This study |
pDML1260 | pDML1251 derivative carrying BlaR1 E253A mutation | This study |
pDML1261 | pDML1251 derivative carrying BlaR1 D257A mutation | This study |
pDML1262 | pDML1251 derivative carrying BlaR1 E274A mutation | This study |
pDML1264 | pDML1251 derivative carrying BlaR1 D221A mutation | This study |
pDML1265 | pDML1251 derivative carrying BlaR1 Y272A mutation | This study |
pDML1266 | pDML1251 derivative carrying BlaR1 E213D mutation | This study |
pDML1267 | pDML1251 derivative carrying BlaR1 E213Q mutation | This study |
pDML1268 | pDML995 derivative carrying BlaR1 H212A/E213A/H216A mutation | This study |
pDML1269 | pDML995 derivative carrying BlaR1 E213A mutation | This study |
pDML1270 | pDML995 derivative carrying BlaR1 H212A/H216A mutation | This study |
pDML1271 | pDML995 derivative carrying BlaR1 H216A mutation | This study |
pDML1273 | pDML995 derivative carrying BlaR1 E253A mutation | This study |
pDML1274 | pDML995 derivative carrying BlaR1 D257Amutation | This study |
pDML1275 | pDML995 derivative carrying BlaR1 E274A mutation | This study |
pDML1277 | pDML995 derivative carrying BlaR1 D221A mutation | This study |
pDML1278 | pDML995 derivative carrying BlaR1 Y272A mutation | This study |
pDML1279 | pDML995 derivative carrying BlaR1 E213D mutation | This study |
pDML1280 | pDML995 derivative carrying BlaR1 E213Q mutation | This study |
pDML3045 | pDML995 derivative carrying BlaR1 R304A/R305A mutation | This study |
pDML1283 | pCR-ScriptTMSK(+) derivative carrying BlaR1 L3 loop coding sequence | This study |
pDML1284 | pDML1283 derivative carrying BlaR1 L3 E213A mutation | This study |
pDML1285 | pDML1283 derivative carrying BlaR1 L3 H212A/H216A mutation | This study |
pDML1286 | pDML1283 derivative carrying BlaR1 L3 D257A mutation | This study |
pDML1287 | pDML1283 derivative carrying BlaR1 L3 D221A mutation | This study |
pDML1293 | pDML1283 derivative carrying BlaR1 L3 E253A mutation | This study |
pDML1288 | pLEX derivative allowing the expression of BlaR1 L3 loop | This study |
pDML1289 | pLEX derivative allowing the expression of BlaR1 L3 carrying E213A mutation | This study |
pDML1290 | pLEX derivative allowing the expression of BlaR1 L3 carrying H212A/H216A mutation | This study |
pDML1291 | pLEX derivative allowing the expression of BlaR1 L3 carrying D257A mutation | This study |
pDML1292 | pLEX derivative allowing the expression of BlaR1 L3 carrying D221A mutation | This study |
pDML1294 | pLEX derivative allowing the expression of BlaR1 L3 carrying E253A mutation | This study |
Apr: ampicillin resistance.
Cmr: chloramphenicol resistance.