Table 1.
Primers used for gene deletion and complementation.
| Gene | Primer Name | Sequence (5’- 3’) | Notes |
|---|---|---|---|
| MrUBI4 | MrUBI4-5F | GGAATTCGAGCAAGACAAGCCAACG, EcoRI | For construction of gene disruption vector |
| MrUBI4-5R | AACTGCAGAAGAAAGCAGGGTCAAGAT, PstI | ||
| MrUBI4-3F | GCTCTAGACAGTAGTTGATTGGACGATG, XbaI | ||
| MrUBI4-3R | GCTCTAGACTGGGAGTAAAGTGGAAGAT, XbaI | ||
| MrUBI4-upF(P5) | GTGGCTGTCATCAGGAGTTT | PCR identification of MrUBI4 deletion transformants | |
| MrUBI4-upR(P6) | GGCATTCATTGTTGACCTCC | ||
| MrUBI4-dnF(P7) | GTTTCTGGCAGCTGGACTTC | ||
| MrUBI4-dnR(P8) | AGCGTGGACAGACTTTGATTT | ||
| MrUBI4CP-5F | GGACTAGTGGGTGGACTGGAGGTA, SpeI | For gene complementation | |
| MrUBI4CP-3R | GCTCTAGATAGGAATCGAACGCAGTT, XbaI | ||
| MrUBI4-F(P1) | GGAAGTCACTAACAATCCCACG | Genomic PCR and RT-PCR analysis | |
| MrUBI4-R(P2) | AAGTCGCAGGACAAGGTG | ||
| bar | bar-F(P3) | TCGTCAACCACTACATCGAGAC | Genomic PCR analysis |
| bar-R(P4) | GAAGTCCAGCTGCCAGAAAC | ||
| ben | ben-F | GGTAACTCCACCGCCATCCA | Genomic PCR analysis |
| ben-R | GCAGGGTATTGCCTTTGGACTT | ||
| gpd | gpd-F | GACTGCCCGCATTGAGAAG | RT-PCR analysis |
| gpd-R | AGATGGAGGAGTTGGTGTTG | ||
| UBI4-probe-F | TGATAAGGACGGCGGTTTG | For probe synthesis | |
| UBI4-probe-R | CCGAAGTAGGCAGCACGAT |
Notes: Gray underlined boxes indicate the restriction enzyme sites.