TABLE 4.
Primers used in this study
| Primer | Sequence (5′–3′)a | Purpose |
|---|---|---|
| pMDpcaA1-2F | TAGAAGGAGACGTAGATGAAAGGACAATGCGACATGG | Forward primer to amplify pcaA1 and pcaA2 |
| pMDpcaA1-2R | CTGCAGGTCGACGATTCAAAGCGGATAGGCGAGGTAG | Reverse primer to amplify pcaA1 and pcaA2 |
| pMDpcaA3F | GGATCCTCTAGAGATATGACCAGGAAGACCTTTGCGA | Forward primer to amplify pcaA3 |
| pMDpcaA3R | TCGAAATCTCCTTGTTTCAGCGTTTCATATATTTCTGGATGTT | Reverse primer to amplify pcaA3 |
| pMDpcaA4F | AACAAGGAGATTTCGATGGCGGAAAAGATTCGGGT | Forward primer to amplify pcaA4 |
| pMDpcaA4R | CTACGTCTCCTTCTATCATTCGGGATCGGGAATGA | Reverse primer to amplify pcaA4 |
| pMDpcaB1-2F | TAGAAGGAGACGTAGTCAGGGGCGACCAAATCACT | Forward primer to amplify pcaB1 and pcaB2 |
| pMDpcaB1-2R | CTGCAGGTCGACGATGCTGTTCCACCGAGCCCAC | Reverse primer to amplify pcaB1 and pcaB2 |
| pETpcaA1-2F | AGGCACCATATGAAAGGACAATGCGACATGG | Forward primer to amplify pcaA1 and pcaA2 with an NdeI site |
| pETpcaA1-2R | TTTAAGCTTTCAAAGCGGATAGGCGAGGTA | Reverse primer to amplify pcaA1 and pcaA2 with an XhoI site |
| pETpcaA3F | AGGCACCATATGACCAGGAAGACCTTTGCG | Forward primer to amplify pcaA3 with an NdeI site |
| pETpcaA3R | TTTAAGCTTTCAGCGTTTCATATATTTCTGGATG | Reverse primer to amplify pcaA3 with an XhoI site |
| pETpcaA4F | AGGCACCATATGGCGGAAAAGATTCGGGT | Forward primer to amplify pcaA4 with an NdeI site |
| pETpcaA4R | TTTAAGCTTGCACGGTGCAGGTGGAAATC | Reverse primer to amplify pcaA4 with an XhoI site |
| pBBR2SF | GGGAACAAAAGCTGGCGCTCACTGCCCGCTTTC | Forward primer to amplify Ptrc |
| pBBR2SR | TGGCGGCCGCTCTAGTCTGCGGACTGGCTTTCTACG | Reverse primer to amplify Ptrc |
| pBR2SHF | TTTAAGCTTAGCGGATAACAATTCCCCTCTAGA | Forward primer to amplify pcaA1 and pcaA2 from pETpcaA1A2 with a HindIII site |
| pBR2SHR | CCGCTCGAGTCAAAGCGGATAGGCGAGGTA | Reverse primer to amplify pcaA1 and pcaA2 from pETpcaA1A2 with an XhoI site |
| pBBRpcaA1-2CF | AAGGAGATATACATACCTTAGCCGCTTGACACCAGA | Forward primer to amplify pcaA1 and pcaA2 |
| pBBRpcaA1-2CR | TCGCTACTCGCCATATTGGTCTGTCATCATTCAAAGCG | Reverse primer to amplify pcaA1 and pcaA2 |
| pBBRpcaB1-2CF | GGGAACAAAAGCTGGTCAGGGGCGACCAAATCACT | Forward primer to amplify pcaB1 and pcaB2 |
| pBBRpcaB1-2CR | TGGCGGCCGCTCTAGGCTGTTCCACCGAGCCCAC | Reverse primer to amplify pcaB1 and pcaB2 |
| pcapF | CAAAAGCTTCGAATTCCATGACGCGGGTCTTTCCG | Forward primer to amplify promoter of pcaA1A2 with an EcoRI site |
| pcapR | CGAAGCTTGGCTGCAGTGCCATGTCGCATTGTCCTTT | Reverse primer to amplify promoter of pcaA1A2 with an PstI site |
| pETpcaCF | AGGCACCATATGATGACAGACCAACAGTCGAATAT | Forward primer to amplify pcaA3 with an NdeI site |
| pETpcaCR | TTTAAGCTTTCAAATTCCTTCGCGTGGAGA | Reverse primer to amplify pcaA3 with an XhoI site |
a
Underlined bases indicate that they overlapped and were used to construct plasmids by the In-Fusion method. The bold bases indicate restriction enzyme sequences.