TABLE 1.
Sequences of primers used
| Category and gene | Primer name | Sequence (5′ to 3′) | Notes |
|---|---|---|---|
| Gene replacement | |||
| Mrdcl1 | Mrdcl1-5F | CGGAATTCGCGAAACAACAGCCAGAGTA | |
| Mrdcl1-5R | CGGGATCCGCAAACAACCCAGACCAGA | ||
| Mrdcl1-3F | GGACTAGTTCGATTTAGTGGGAATGAGC | ||
| Mrdcl1-3R | GCTCTAGACATCGCTTGAATCTCCTGTG | ||
| Mrdcl1-NF | TTGATTGCGATTCGGGTAG | PCR identification of Mrdcl1 deletion transformants | |
| Mrdcl1-NR | CGTCAACATCCTCCTCACC | ||
| Mrdcl2 | Mrdcl2-5F | AACTGCAGCAAAGGTTCCTCGCCAGTCC | |
| Mrdcl2-5R | AACTGCAGATCCCGCCATTCAACTAACG | ||
| Mrdcl2-3F | GGACTAGTCAGAAACAGCCGTTACATCC | ||
| Mrdcl2-3R | GCTCTAGAGCTCTGCCATTATTCTTGTCG | ||
| Mrdcl2-NF | CCGACACCTGGGATACGAT | PCR identification of Mrdcl2 deletion transformants | |
| Mrdcl2-NR | AGTTGCTCGCTCCTTTACG | ||
| Mrago1 | Mrago1-5F | CGGAATTCATCGCTTCCCGACTGTGA | |
| Mrago1-5R | CGGGATCCTGGCCGCTGCTGCTAAAT | ||
| Mrago1-3F | GGACTAGTTGGTCTCGTCGCTTCTGT | ||
| Mrago1-3R | GCTCTAGATGCTTGGGCATCGACTTG | ||
| Mrago1-NF | TCCCGAGACACTGATAGGC | PCR identification of Mrago1 deletion transformants | |
| Mrago1-NR | CAACGACTTATGGACCTG | ||
| Mrago2 | Mrago2-5F | AACTGCAGTTGACCTCGGTGCTTTCG | |
| Mrago2-5R | AACTGCAGGGCGCTGAGGGTTGCTAT | ||
| Mrago2-3F | GGACTAGTAGTCGGATATTGTGAAAGCG | ||
| Mrago2-3R | GCTCTAGACCTTTGGCTTGTATGTCTGC | ||
| Mrago2-NF | ATGTCTAGTGGGCATAGAGGTG | PCR identification of Mrago2 deletion transformants | |
| Mrago2-NR | AGCTGCTGAGCTGTCTGG | ||
| BAR | bar-F | GGAGGTCAACAATGAATGCC | |
| bar-R | CCACGTCATGCCAGTTCC | ||
| BEN | ben-F | ATGGCTACCTACTCCGTCGTG | |
| ben-R | CTCGTCCATACCCTCACCA | ||
| Gene complementation | |||
| Mrdcl1 | Mrdcl1cp5F | GCTCTAGAGCGAAACAACAGCCAGAG | |
| Mrdcl1cp3R | GCTCTAGATCCCGTCGTGTTCTTCTT | ||
| Mrdcl2 | Mrdcl2cp5F | GCTCTAGAGCCAAGGCAGTGAAGTAT | |
| Mrdcl2cp3R | GCTCTAGACTGCCATTATTCTTGTCG | ||
| Mrago1 | Mrago1cp5F | GCTCTAGAGACGAAAGAGGCATACACT | |
| Mrago1cp3R | GCTCTAGACTGCTTGGGCATCGACTT | ||
| Mrago2 | Mrago2cp5F | GCTCTAGAGGTCACCAAATTGCACCTA | |
| Mrago2cp3R | GCTCTAGAGAGCCACTGCCAGTATAAGT | ||
| Gene expression analysis and RT-PCR identification of deletion transformants | |||
| Mrdcl1 | Mrdcl1F | GAAGGCGGTCACTGTAATGG | |
| Mrdcl1R | ATGGGTATGACGGATGCTTG | ||
| Mrdcl2 | Mrdcl2F | CCGACTTCCTTCATCCTGTT | |
| Mrdcl2R | TTCGCAATCAATGTGACCTC | ||
| Mrago1 | Mrago1F | GAGAAGACCGAAGTAAAGGATG | |
| Mrago1R | CGGACGGAATGTTGATAGTG | ||
| Mrago2 | Mrago2F | GCATCAATCAGACGGTGGAG | |
| Mrago2R | ATTAGAGGAGGAGCCAGGTGAG | ||
| gpd | gpdF | GACTGCCCGCATTGAGAAG | |
| gpdR | AGATGGAGGAGTTGGTGTTG |