Table 2.
Primers used in the construction and confirmation of recombinant pCT plasmids
| pCT gene | Description | Primer sequence | Amplimer size (bp) (WT/inactivation) |
|---|---|---|---|
| Sigma factor pCT_066 |
Confirmation forward |
ACAGCGTCTTCTCGTATCCA |
1289/1675 |
| Confirmation reverse |
GTTCTTCCAGCTGACGTAAC |
1289/1675 |
|
| Recombination 1 |
GGAGGGCGTCTCGCTAAAAAAACTTACTCAAACACATCAAGTGTAGGCTGGAGCTGCTTC |
1574 |
|
| Recombination 2 |
GCATTACTTTTTATTCTCGTGAGACTCAAGGTCATTCGGTGGGAATTAGCCATGGTCCAT |
1574 |
|
|
rci |
Confirmation forward |
AAGGTCATCTGCAGGAGT |
945/1867 |
| Confirmation reverse |
GTGTCGCAGCAACAATA |
945/1867 |
|
| Recombination 1 |
GGGGGACATGCCGTATGAATCCTGTTGAACTGGTCCGAAAGTGTAGGCTGGAGCTGCTTC |
1574 |
|
| Recombination 2 |
GCAGTGTCACGACAAACAGCCCGTTTCTGCACCCGACAGTGGGAATTAGCCATGGTCCAT |
1574 |
|
|
pilS |
Confirmation forward |
GCGGAAGGAAGTGAGCATAA |
722/2053 |
| Confirmation reverse |
CAGTGACATGCTGAAGCAGT |
722/2053 |
|
| Recombination 1 |
TGGTGACCAGATCAATACAGTTTTTCTTCGGCACATTGCTGTGTAGGCTGGAGCTGCTTC |
1574 |
|
| Recombination 2 |
AACCTGCAGACAATCGCCACCAAAATGAAAGCCCAGAAAGGGAATTAGCCATGGTCCAT |
1574 |
|
|
traY |
Confirmation forward |
GGAGAGTCCGGTCTGTATGA |
2423/2138 |
| Confirmation reverse |
TGCAACCAGTGTGGTACAG |
2423/2138 |
|
| Recombination 1 |
GTATCCTGGTCTGCCTGTTACTGATGAGTACCATTGCAGCGTGTAGGCTGGAGCTGCTTC |
1574 |
|
| Recombination 2 |
CGGCACAAAACAGCAAAAACAGCAGGAAGTAGAGTGGTGGGGGAATTAGCCATGGTCCAT |
1574 |
|
|
pndACB |
Confirmation forward |
AAGGATTGTGGCGGACAGGA |
486/1288 |
| Confirmation reverse |
TGATGACGCACAGGACGGAA |
486/1288 |
|
| Recombination 1 |
CCCAGGCGATTTTTTTATCAATCAACCCAGGGCCCACTGTGTGTAGGCTGGAGCTGCTTC |
1574 |
|
| Recombination 2 |
ATTGAGGTCAGCCTTCGCAACAATCCGGCGGCAGATGTCCGGGAATTAGCCATGGTCCAT |
1574 |
|
| parB | Confirmation forward |
TATTAAAAATAACGCGGCGG |
663/1872 |
| Confirmation reverse |
GCAAAGTATCACACTGCCAAAA |
663/1872 |
|
| Recombination 1 |
GGAGCGGCGGGAGAGTATAGTCATTATTGTAGTCCGGGTAGTGTAGGCTGGAGCTGCTTC |
1574 |
|
| Recombination 2 | CTTTTCACTCACCATTATTTTTTCCGCTTCTCTCTGTGCCGGGAATTAGCCATGGTCCAT | 1574 |