TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Genotype or relevant characteristic | Reference or source |
|---|---|---|
| Strains | ||
| E. coli BL21[λDE] CodonPlus | F−ompT hsdSB (rB− mB−) dcm met gal(λDE3)endA Hte[argU ileY leuW Tetr] | Stratagene |
| E. coli DH5α | K-12 φ80d lacZΔM15 endAI hsdR17 (rK− mK−) supE44 thi-1 gyrA96 relA1 Δ(lacZYA-argF+)U169 F− | Invitrogen |
| E. coli W3110 | F−trpA23 xyl glySL | 16 |
| E. coli MHD79 | sltY mltA mltB mltC mltD emtA (Cmr Kanr Tetr) | 15 |
| E. coli MC1061 | araD139 Δ(ara-leu)7696 Δ(lac)174 galU galK hsdR2(rK− mK−) mcrB1 rpsL (Strr) | 4 |
| Pseudomonas aeruginosa PAO1 | Serotype O5; A+ B+ | 13 |
| Plasmids | ||
| pET30a(+) | IPTG-inducible expression vector; Kanr | Novagen |
| pET32-EK/LIC | IPTG-inducible expression vector; Ampr | |
| pET30a(+)-Ampr | pET30a(+) derivative containing bla on a DraIII/AlwNI fragment; Ampr | This study |
| pACJW2 | pET30a(+) derivative containing P. aeruginosa pbpA on an NdeI/HindIII fragment; Kanr | 23 |
| pACBL17 | pACJW2 derivative encoding the N-terminal n-PB module of P. aeruginosa PBP 2 of Met1to Ser250; Kanr | This study |
| pACBL18 | pET30a(+) derivative containing E. coli pbpA on an NdeI/HindIII fragment; Kanr | This study |
| pACBL19 | pACBL18 derivative encoding site-specific replacement of Ser330→Ala on E. coli pbpA; Kanr | This study |
| pACBL24 | pACBL18 derivative encoding the N-terminal n-PB module of E. coli PBP 2 of Met1 to Thr252; Kanr | This study |
| pACBL26 | pET30a(+)-Ampr derivative containing mutant E. coli pbpA on an NdeI/HindIII fragment encoding site-specific replacement of Ser330→Ala; Ampr | This study |
| pACBL28 | pET30a(+)-Ampr derivative containing mutant E. coli pbpA on an NdeI/HindIII fragment encoding N-terminal n-PB module of Met1 to Ser253; Ampr | This study |
| pACBL29 | pET30a(+)-Ampr derivative containing E. coli pbpA on an NdeI/HindIII fragment; Ampr | This study |
| pACCA11 | pET30a(+)-Ampr derivative containing P. aeruginosa pbpA on an NdeI/HindIII fragment; Ampr | This study |
| pACDC38 | pACJW2 derivative encoding site-specific replacement of Ser327→Ala on P. aeruginosa pbpA; Kanr | 23 |
| pACKD16 | pET30a(+) derivative containing pbpA on an NdeI/HindIII fragment encoding an N-terminal truncation of PBP 2 of Pro2 to Arg38; Kanr | 23 |
| pACSR1 | pET30a(+) derivative containing P. aeruginosa ftsI on an NdeI/XhoI fragment encoding PBP 3; Kanr | This study |
| pUCH6-wzz1 | wzz1 cloned via XbaI-PstI from pETH6-wzz1 into pUCP27 (N-terminal His6 tag) | 6 |