TABLE 2.
Oligonucleotides used in this study
| Description and applicationa | Sequence (5′→3′)b | Created restriction site | Primer |
|---|---|---|---|
| Constructions | |||
| pGID121 | AAAGCTTCGCGTTTTTCGTCATGATT | HindIII | C13 |
| ATCTAGACATAAATTCATCCCCATTCT | XbaI | A12 | |
| pGID118 | AGCTAGCAACAGGAGATCCGTTTGAG | NheI | A15 |
| CTTACCATAAAATTCCACTT | E2 | ||
| pGID112 | CAAAGCTTTTACTGCAAACA | HindIII | D1 |
| AAGAATCCGCAACTTTCATGG | D2 | ||
| pGID109 | CTATGAGTCTAGATTGCTTCATG | XbaI | D3 |
| AACAAATTCGCCAACATTCC | D4 | ||
| RNA analysis | |||
| qRT-PCR drm | GCGGAAAAACCACTTGCTTACT | DRMF2 | |
| CGGAAAGGTGTGTCAATGTATAGG | DRMR2 | ||
| qRT-PCR agrB | CGGCAGACACAGAAAGTTTG | BF2 | |
| TGCGAATGGTATTAGCAACG | BR2 | ||
| qRT-PCR agrD | AAATCAGTTGGTAAATTCCTTTCTAG | DF2 | |
| AATGGACTTTTTGGTTCGTATACA | DR2 | ||
| qRT-PCR agrC | GGGGTCAATCGCAGGTTTTG | CF2 | |
| CTTTAAGTTCGTTGGTTGCCGTA | CR2 | ||
| qRT-PCR agrA | GCAAGCAGAAGAACGATTTCCAA | AF2 | |
| CGCTGTCTCAAAAAACAAGATAT | AR2 | ||
| RT-PCR agrBD | CAAAGCTTTTACTGCAAACA | HindIII | D1 |
| TTGCATTTTCACAAATGGACTT | DR | ||
| RT-PCR agrDC | CCATGAAAGTTGCGGATTCT | DF | |
| GGCACTTACAAAAACAATCAATAC | C35 | ||
| RT-PCR agrC and Northern blot agrC | GGAATGTTGGCGAATTTGTT | C11 | |
| CTTCAAACCCGGCATATCAT | C10 | ||
| RT-PCR agrCA | CAAAAGGAGAAGGTCGTGGA | C23 | |
| ACCATTCATGTCCGGCTGCC | A46 | ||
| Northern blot agrB | ACCATGGGTAAATTCGTTGTAAAATA | NcoI | B17 |
| CGTGCAATTCAATGTTTTGG | BR | ||
| Northern blot agrA | AGCTAGCAACAGGAGATCCGTTTGAG | NheI | A15 |
| CTTACCATAAAATTCCACTT | E2 | ||
| 5′RACE agrB | TGCGAATGGTATTAGCAACG | BR2 | |
| CGAAATCAACACATCCGCCAT | B34 | ||
| 5′RACE agrC | GGCACTTACAAAAACAATCAATAC | C35 | |
| CTAAAGTAAATAAAGGGAAGGCTA | C36 | ||
| 5′RACE agrA | TTGATGTGTAGGCATTCGTG | A44 | |
| ACCATTCATGTCCGGCTGCC | A46 | ||
| CGCTGCATTCTGTTATCTTCA | A48 | ||
| CTTCAATATATTTCGTTAACCT | A50 | ||
| Primer extension agrB | GACAAAGGGACTTTTGCAGT | B18 | |
| CTTCTTCATCATCTTTCCAGCG | B26 | ||
| CGAAATCAACACATCCGCCAT | B34 |
a
qRT-PCR, quantitative RT-PCR.
b
Specific restriction sites are underlined, and extra nucleotides added to include restriction sites to the PCR product are indicated in boldface.