Table 2. Oligonucleotides used in this study.
Oligonucleotide Primers | Sequence (5′-3′)a |
Recombinant expression of GST-Har and variants | |
HarNterm | CGGAATTCATCGAAGGTCGTATGATAGTCACATCA |
HarCterm | CGCTCGAGCCATATCGGATCAATGTTATATGTCT |
Har150_Nterm | CCGCGTGGATCCATGATAGTCACATCACTG |
Har150_Cterm | CGTGGATCCCGGGTTACTGCTTCTTCCTGCATTT |
HarC97A SDM | GCTTCATTTGCAGAGCAGGCACCGCTGCTTTTCTGTACC |
DNA target for EMSA | |
PGN_0001_240bp_For | GGTGTTGATAACTCGGTCGCGCCTT |
PGN_0001Rev | CTAAAAAAATATCGTTTTGAGAGCAGT |
Construction of har mutant | |
PGN_1502-Fwd | ATGTCGCCTTCCGAGGCTAT |
ErmF-PGN_1502-Rev | GCAATAGCGGAAGCTATCGGTTATCTTTTCGATCCATTCTTGC |
PGN_1502-ErmF-Fwd | AGAATGGATCGAAAAGATAACCGATAGCTTCCGCTATTGC |
Term-ErmF-Rev | GTCTTTCGACTGAGCCTTTCGTTTTAGCATCTAATTTAACTTCAATTCC |
Term-Prom-region-Fwd | GCTCAGTCGAAAGACTGGGCCTTTCGTTTTACGGAGTGAAAAAGGAGCCG |
PGN_1504-Prom-region-Rev | CAATGTTATATGTCTGTGTTATCTCTCTTTTACATCATATTTTCC |
Prom-region-PGN_1504-Fwd | AATATGATGTAAAAGAGAGATAACACAGACATATAACATTGATCC |
PGN_1504-Rev | GCAGATATTTTGTAGCCTCCATC |
Construction of har complement | |
PGN_1502-Fwd | ATGTCGCCTTCCGAGGCTAT |
CepA-Har-Rev | ACTTTCCTTAACTCTTTTGACGTCTTATTTTTTCTTCTTGGGAGCGGCT |
Har-CepA-Fwd | AGCCGCTCCCAAGAAGAAAAAATAAGACGTCAAAAGAGTTAAGGAAAGT |
ErmF-CepA-Rev | TGTGTAGGTTCTAATTGAAGGACAGACGTCTCAAGTCACCGATAG |
CepA-ErmF-Fwd | CTATCGGTGACTTGAGACGTCTGTCCTTCAATTAGAACCTACACA |
ErmF-Rev | GATACTGCACTATCAACACACTC |
qRT-PCR | |
PGN_0287 RT Fwd | CCAGCAGCACTTTCCATACAAA |
PGN_0287 RT Rev | CCACTGATTACGGCCTCATTT |
PGN_0448 RT Fwd | AGTAAAGGGGTAGGGCAACG |
PGN_0448 RT Rev | ATCGGATTCGTGTTCCAAAGC |
PGN_1296 RT Fwd | CCTGCAAGAGCGTGAAGTTG |
PGN_1296 RT Rev | GGATCGGAAAGCCGTATAAGC |
PGN_1578 RT Fwd | TGTTGTGGAAAGGAGTGTGG |
PGN_1578 RT Rev | AGAAGGAATGAAGTCGGTTGTT |
PGN_2083 RT Fwd | GCATTCTTTTCTGGCGTAGCA |
PGN_2083 RT Rev | TTTGCGAAACGGCACTCCCT |
Underlined sequence of SOE primers indicates the part of the primer that is complementary to the target sequence, with the remainder of the primer providing complementarity with a second PCR product for splicing.