Table 1.
Oligonucleotide sequences.
| Name | Sequence 5′–3′ | Description |
|---|---|---|
| chiA intron deletion | ||
| Exon2Fwd | ACGCACGAGACCGGACATTTCTGCTACATCAGC | Exon2 forward primer |
| Exon1Rev | GATGTAGCAGAAATGTCCGGTCTCGTGCGTGAC | Exon1 reverse primer |
| Exon1Fwd | TCAACGGCATCAAGAACCAG | Exon1 forward primer |
| Exon2Rev | GTCTTGAACGCGATCACGGC | Exon2 reverse primer |
| ChitA amino acid mutagenesis | ||
| rChitA‐B73(A47D)Fwd | GCGGCACGACCGACGACTACTGCGGCGACGGGT | pTAN112ΔI(A47D) |
| rChitA‐B73(A47D)Rev | CGTCGCCGCAGTAGTCGTCGGTCGTGCCGCAGT | pTAN112ΔI(A47D) |
| rChitA‐LH82(D48A)Fwd | GCGGCACGACCGACGCCTACTGCGGCGACGGGT | pTAN114ΔI(D48A) |
| rChitA‐LH82(D48A)Rev | CGTCGCCGCAGTAGGCGTCGGTCGTGCCGCAGT | pTAN114ΔI(D48A) |
| FwdMut | ATGGCAAACGCGCCGAGGATCCTG | Both mutants |
| RevMut | CGATCTCGCGCTTGCCCTCCACCT | Both mutants |
| Expression plasmid cloning | ||
| LH82ExpFwd | GGATGAATTCCAGAACTGCGGCTGCCAGCCTAAC | pTAN121 and pTAN121(D48A) cloning |
| B73ExpFwd | GGATGAATTCCAGAACTGCGGCTGCCAGCCAAAC | pTAN122 and pTAN122(A47D) cloning |
| ExpRev | CCATTCTAGACTAGCAAGTGAGGTTGGGCCCTGG | All four expression clones |