TABLE 1.
Primers used in this study
| Primera | Target gene | Sequence (5′-3′)b |
|---|---|---|
| For gene deletion | ||
| CgCNB 100-F | CNB1 | GTATGTGATGCTTCTCACAGGGTTCAGACGGTTACATACCATCGCTTGAGAGTCATAGTAAATGTTCAGGTTCACGATTAAATCATGCTTTCTCTTTGATAATACGACTCACTATAGGGC |
| CgCNB 100-R | CNB1 | GCGAACTCTGAAATGTAGATCAAGGATTATTCTGTCCTTGAAATGGGTGTTGATGTCCCTCACTAGGAAAGACAACCACTTTACTATTGTAAGGGGTGACGCTCTAGAACTAGTGGATCC |
| CgCRZ 100-F | CRZ1 | GATAACGAGTTGGACGCCCTCTTTTGGAAGTCTGTTCTGGTTGCAGATGCTTATAGACCCTGGATCAAGCACTTCATTTCATTGGGATTACAGCTTTTCTAATACGACTCACTATAGGGC |
| CgCRZ 100-R | CRZ1 | CACAATCTTGATTCTGAAGAAAAAAATTTATCATTAAAAATACTGGAGGTTTGTGTTAATTTATTCCAAAGTAACACCCATCTCAGTTGCTTGAATATTCGCTCTAGAACTAGTGGATCC |
| For gene cloning | ||
| CgCNB1-F2-5P | CNB1 | ATCAAGGGAAATGGGAGC |
| CgCNB1-R1-5P | CNB1 | CGCCCTAAGTTACATCTCTCCTCG |
| CgCRZ1-F1-E | CRZ1 | CGGAATTCATGGGCGATAACGAAGAGGA |
| CgCRZ1-R1938-E | CRZ1 | CGGAATTCTTATTCCAAAGTAACACCCATCTCA |
a
“F” and “R” indicate forward and reverse primers, respectively.
b
Sequences homologous to flanking regions of the target ORF are shown in italics. Sequences shown in boldface are present in pBSK-HIS. Restriction sites are underlined.