Table 1.
Primers and probes of Shk1 used in this study
| Primer code* | Sequence (5′→3′)† | Relevant characteristic |
|---|---|---|
| P1 | ATGGGGGACACTACGATAGC | Amplify the full cDNA sequence of the Shk1 gene |
| P2 | CAGAACTGCTTAGTACAGGTT | |
| P3 | AAAAGGTTGAACGGGCTTGTGTC | Amplify the left homologous arm of the Shk1 gene of S. sclerotiorum (1450 bp) |
| P4 | CCACCAGCCAGCCAACAGCTCCCATCGCAGTCGTGTGAGCTATCG | |
| P5 | CAATACGCAAACCGCCTCTCCCCGTCCTTCACTTGTAACGGCAG | Amplify the right homologous arm of the Shk1 gene of S. sclerotiorum (1541 bp) |
| P6 | ATGACATCTTGCGGACTTGGGGC | |
| P7 | GGGAGCTGTTGGCTGGCTGGTGG | Amplify the hph gene (1764 bp) |
| P8 | GGGGAGAGGCGGTTTGCGTATTG | |
| P9 | AGAACACTGACTGATGAAAGG | Amplify the knockout vector of the Shk1 gene of S. sclerotiorum (4260 bp) |
| P10 | CAGGGTGAAAGATAGGATACT | |
| P11 | AACGGAGGGTATTCTCGGGG | Amplify a partial fragment of the Shk1 gene of S. sclerotiorum (482 bp) |
| P12 | TGTCTTTCAGCGAAGCGATT | |
| P13 | CAAAGCATCAGCTCATCGAGAG | Amplify a partial fragment of the hph gene (503 bp) |
| P14 | GAAAAGTTCGACAGCGTCTCC | |
| P15 | GAGTAAAAACCGTTGATG | Confirm whether the hph genes homologously replaced the Shk1 gene of S. sclerotiorum (2255 bp) |
| P16 | GTCTGGACCGATGGCTGT | |
| P17 | TCATTTGGATGCTTGGGTAG | Confirm whether the hph genes homologously replaced the Shk1 gene of S. sclerotiorum (2333 bp) |
| P18 | CCTTAAACCAGATTCCGTAC | |
| P19 | CTGCAGAAccaccatgttggGTCGACAGAAGATGATATTG | Amplify the NEO cassette containing a trpC promoter (1181 bp) |
| P20 | CCGctcgagTCAGAAGAACTCGTCAAGAAGGCG | |
| P21 | TAAgcggccgcGCTGTACGAGTGATGCGA | Amplify the Shk1 gene (include the control region of the Shk1 gene) (6598 bp) |
| P22 | TAActgcagATAGTAGCAAACACTGCCAG | |
| P23 | TGAAGGGAAAGAGGAGAAGA | Amplify a probe for Southern blotting (560 bp) |
| P24 | GATTGGGAGCAGGATAAGAA | |
| P25 | TCAAGCCAAGCAACATCCTCGT | Amplify the SsHOG1 gene for quantitative real‐time PCR |
| P26 | ATCCCGCACTCCAAACATCAAC | |
| P27 | GAGTTTGTCGGTGCTCCCTTTG | Amplify the PAK gene for quantitative real‐time PCR |
| P28 | ATCATACGCTGCCATTCCTTCG | |
| P29 | CCCCAGCGTTCTACGTCT | Amplify the reference gene actin for quantitative real‐time PCR |
| P30 | CATGTCAACACGAGCAATG |
*The number code refers to the primer binding sites shown in Fig. 1A.
†The respective restriction enzyme sites included in the primers are listed in lowercase letters in the sequence.