Table 4 Oligonucleotide sequences used in this study
Primer | Sequence (5`→ 3`) | Amplicon (bp) | Target gene |
|---|---|---|---|
ldi deletion construct | |||
(pK19mobsacB | |||
ORF25_Eco RI_F | TCGTAGAATTC CATGCCTGCGCACGCTGATG | 1307 | ORF25 |
ORF25_Xho I_ATG_R | GAGACTCGAG ATGTTCAGTCGCATGTCGTCT | ||
ORF27_Xho I_TAA_F | TATACTCGAG TAAGGGGGACGCGGCGGCCTG | 763 | ORF27 |
ORF27_Hind III_R | TCGTAAAGCTT ATGGACGACGGCACATGGA | ||
p27 + _F | ACGAAGCCGAGCATGCCCAC | 2199 | encompassing |
p27 + _R | AGCAGCAGGCCGACGTGTTC | ORF27 | |
p27mismatch_F | CGCCCGGTTCGAGGAAGG | - | nucleotide |
p27mismatch_R | CCCTTCCTCGAACCGGGCG | exchange | |
geoA deletion construct | |||
(pK19mobsacB) | |||
ORF2930_Xba I_F | TCTAGA CCACCAGGGCGCATGCTTCAGTT | 1749 | ORF2930 |
ORF2930_Xho I_R | CTCGAG TGAGCAGGGCGCGACTCC | ||
ORF32_Xho I_F | CTCGAG CATCGTTGAGTGTCTCCTGGTTG | 1712 | ORF32 |
ORF32_Hind III_R | AAGCTT TGGAAACGACATAGGGGACAGGA | ||
Control of ldi or geoA deletion | |||
ldi _F | CGCCACCACCGAGGACTATTTC | 432 | ldi |
ldi _R | AGGTGGGCATGCTCGGCTTCGTA | ||
ORF25_401_F | GAAGGTGCGCGGCAAGGAATA | 2463a/ | ORF25-27 |
ORF27_2005_R | CATGGACAGCGGCACACGGGCAA | 1269b | |
geoA _260_F | ACCGGGTCGTGCTGTCCTTCAAT | 284 | geoA |
geoA _527_R | CGCGCCGGTCTGGATGC | ||
ORF30_30967_F | CCAGACGCCGCCGATGATGAAGAG | 1904a/ | ORF30-32 |
ORF32_32822_R | TATCTGAACAAGCCCGAACTGACC | 740c | |
ldi complementation construct | |||
(pBBR1MCS-4) | |||
ldi _Eco RI_F | TGCGGAATTC ATGCGGTTCACATTG | 1206 | ldi |
ldi _Bgl II_R | CGCGAGATCT TTATTTCCCTGCGA | ||
geoA complementation construct | |||
(pBBR1MCS-2) | |||
geoA _Xba I_F | AATCTAGA CGCCCTGCTCAGAACAC | 1290 | geoA |
geoA _Hind III_R | GAGCAAGCTT ACCCTGCGCAAGCAGTTC | ||
Control of adjacent gene transcription | |||
ORF25_254_F | CCCACCGGCTTCTCGTAGTC | 535 | ORF25 |
ORF25_788_R | GCAAGGGCCTGGGCGTGATGTC | ||
ORF27_28_F | CATGGACAGCGGCACACGGGCAA | 350 | ORF27 |
ORF27_377_R | CAGTGGACCTCGCCGTGGAAAT | ||
ORF30_315_F | CGCGGGCGGGATGGTGAT | 411 | ORF30 |
ORF30_725_R | CGGGCGGCGGGTTCGTT | ||
ORF32_608_F | CCTGGCGGCCGGACGACAT | 462 | ORF32 |
ORF32_1069_R | CGCCGGAAGGGGAAACGAC |