TABLE 2.
Primers used in this study
| Primer | Primer sequence | Characteristics |
|---|---|---|
| PAMS F002 | 5′-AGG CAC CCC AGG CTT TAC AC-3′ | External sense primer for site-directed mutagenesis of all signal sequence mutants |
| PAMS R002 | 5′-GTA GAG GCG GTT CGG ATT GG-3′ | External antisense primer for site-directed mutagenesis of all signal sequence mutants, antisense primer for RT-PCR of plcHΔss mutant |
| PLCHR | 5′-CTT CAC TGA TCG CCT GGT TC-3′ | Reverse primer for qPCR studies |
| PLCHF | 5′-CAG TTG AAC ATC CGC AAT CTC-3′ | Forward primer for qPCR studies |
| R8K-1 | 5′-TGG AAA TTC AAG CGT CGA ACC-3′ | Sense primer to generate the plcHR8K mutant |
| R8K-2 | 5′-GGT TCG ACG CCT GAA TTT CCA-3′ | Antisense primer to generate the plcHR8K signal sequence mutant |
| R8S-1 | 5′-TGG AAA TTC TCG CGT CGA ACC-3′ | Sense primer to generate the plcHR8S signal sequence mutant |
| R8S-2 | 5′-GGT TCG ACG CGA GAA TTT CCA-3′ | Antisense primer to generate the plcHR8S signal sequence mutant |
| R8A-1 | 5′-TGG AAA TTC GCC CGT CGA ACC-3′ | Sense primer to generate the plcHR8A signal sequence mutant |
| R8A-2 | 5′-GGT TCG ACG GGC GAA TTT CCA-3′ | Antisense primer to generate the plcHR8A signal sequence mutant |
| R10K-1 | 5′-TTC CGC CGT AAG ACC TTT CTC-3′ | Sense primer to generate the plcHR10K signal sequence mutant |
| R10K-2 | 5′-GAG AAA GGT CTT ACG GCG GAA-3′ | Antisense primer to generate the plcHR10K signal sequence mutant |
| R34A-1 | 5′-GAG ACG CTC GCG CGC GCC CTG-3′ | Sense primer to generate the plcHR34A signal sequence mutant |
| R34A-2 | 5′-CAG GGC GCG CGC GAG CGT CTC-3′ | Antisense primer to generate the plcHR34A signal sequence mutant |
| R35A-1 | 5′-ACG CTC CGG GCG GCC CTG GCC-3′ | Sense primer to generate the plcHR35A signal sequence mutant |
| R35A-2 | 5′-GGC CAG GGC CGC CCG GAG CGT-3′ | Antisense primer to generate the plcHR10K signal sequence mutant |
| R3435A-1 | 5′-ACG CTC GCG GCG GCC CTG GCC-3′ | Sense primer to generate the plcHR34AR35A signal sequence mutant |
| R3435A-2 | 5′-GGC CAG GGC CGC CGC GAG CGT-3′ | Antisense primer to generate the plcHR34AR35A signal sequence mutant |
| PlcH ss-1 | 5′-CAT ATG GTC GAG CCG GAC ATC-3′ | Sense primer to generate the plcHΔss mutant (engineered NdeI site is underlined) |
| PlcH ss-2 | 5′-CAT ATG TTT ATT TCC CGG GTG-3′ | Antisense primer to generate the plcHΔss mutant (engineered NdeI site is underlined) |
| PAMS SEQ3 | 5′-ACG AGT TCT CCC CCT ATC AC-3′ | Sense primer for RT-PCR of plcHΔss mutant |
| OmlA-1 | 5′-TGA TAG ACC AGT TGC GTC CCG-3′ | Sense primer for RT-PCR of omlA |
| OmlA-2 | 5′-TGC TGC CCT CCT TGC CGA G-3′ | Antisense primer for RT-PCR of omlA |
| TAT 6518 | 5′-CGG CCT GAA CCT ACA CAT TGC-3′ | Sense primer to confirm Δtat mutant |
| TAT 8931 | 5′-CAT TAC TCC AGG CAG TGC GAC-3′ | Antisense primer to confirm Δtat mutant |