TABLE 1.
Oligonucleotide primers used in this study and their relevant characteristics
| Primer | Sequence (5′→3′) | Relevant characteristics |
|---|---|---|
| CYP51-Deg-F | GCC(C/T)CC(C/T)GT(C/T)GT(T/G)TTCCAC | Degenerate PCR primers for amplification of a |
| CYP51-Deg-R | AGC(A/G)CC(A/G)AA(A/G)GG(A/G)AG(A/G)TA | 1.3-kb CYP51 fragment from B. jaapii |
| RA2 | (A/C/T/G)CAGCT(A/T)(C/G)CT(A/C/T/GT)(C/G)CTT | Arbitrary primers for a TAIL-PCR for amplification |
| RA3 | GT(A/C/T/G)CGA(C/G)(A/T)CA(A/T/C/G)A(A/T)GTT | of upstream and downstream sequences of the |
| RA4 | CA(A/C/T/G)GCT(A/T)(C/G)GT(A/T/C/GT)(C/G)CAA | 1.3-kb CYP51 fragment from B. jaapii |
| TAIL-SP-Upstr-CYP51-1 | AGGTGAAGCAATCGCCGTACT | Specific primers for a TAIL-PCR for amplification |
| TAIL-SP-Upstr-CYP51-2 | CAGAAAGCAACTGACCTTCGC | of upstream sequence of the 1.3-kb CYP51 |
| TAIL-SP-Upstr-CYP51-3 | CTCGAAAAAGAACCTGTACGGG | fragment from B. jaapii |
| TAIL-SP-Downstr-CYP51-1 | ACTCGTCATTCCAACATCGCA | Specific primers for a TAIL-PCR for amplification |
| TAIL-SP-Downstr-CYP51-2 | AGGTTTTGAACAATCAAGTCGAGG | of downstream sequence of the 1.3-kb CYP51 |
| TAIL-SP-Downstr-CYP51-3 | CGACTATGGGTACGGTCTGGTC | fragment from B. jaapii |
| Bj-CYP51-F | GATCTGGCACACGTGGAAAGA | PCR primers for amplification of the complete |
| Bj-CYP51-R | AACACCGTCGGGGTGAGAAT | CYP51 gene from B. jaapii |
| Bj-CYP51-5-Upstr-F | AGGACCGGAAGGAGGAGTTGA | PCR primers for amplification of the 472-bp 5′ end |
| Bj-CYP51-5-Upstr-R | TGGCGTTTGGGCAATCGTATA | of CYP51 and 610-bp upstream DNA |
| GeneRacer 5′ primer | CGACTGGAGCACGAGGACACTGA | PCR primers for amplifying 5′-end sequences of |
| RACE-SP | TGTGATGGTAGACCCAACGA | cDNA of CYP51 |
| GeneRacer 5′ nested primer | GGACACTGACATGGACTGAAGGAGTA | Nested PCR primers for amplifying 5′-end sequences |
| RACE-nested-SP | TGGCTTAGCACATTGAGCAC | of cDNA of CYP51 |
| Det-Poly-F | AGCCCATCCTTGTGCTGACTT | PCR primers for detecting polymorphic upstream |
| Det-poly-R | TTGCAGACGCGACACGACT | sequences of the CYP51 genes from fenbuconazole-resistant B. jaapii isolates |
| Bj-CYP51-RT-F | CAAAGGTCAATCAGGAACTGCAGA | Real-time PCR primers for determining expression |
| Bj-CYP51-RT-R | AAGAGGAGCCCAGTGAAGCATAAA | level of the CYP51 gene in B. jaapii |
| Bj-Tub-RT-F2 | TCGTCCCATCTCCCAAGGTTT | Real-time PCR primers for determining expression |
| Bj-Tub-RT-R2 | AGACCAAGTGGTTGAGATCTCCG | level of β-tubulin gene in B. jaapii |