TABLE 2.
Plasmids and oligonucleotides used
| Name | Description | Source (reference) or sequencea |
|---|---|---|
| Plasmids | ||
| pBS-KS(+) | Plasmid vector used for cloning purpose | Stratagene |
| pACT1 | Plasmid harboring ACT1 gene | Gift from Anand Bachhawat |
| pSAT1 | Plasmid harboring caSAT1 marker | Reuss et al. (33) |
| pLACZ6 | Plasmid harboring lacZ reporter gene | Gaur et al. (11) |
| pADH1G3 | Plasmid harboring GFP reporter gene and caSAT1 marker | Hiller et al. (14) |
| pCPG1 | Plasmid harboring PCDR1-GFP fusion for ACT1 locus integration | Gaur et al. (11) |
| pCPG3 | Plasmid harboring PCDR1-GFP fusion for CDR1 locus integration | This study |
| pCPG2 | Plasmid harboring CDR1-GFP fusion for CDR1 locus integration | Prasad et al. (30) |
| pCPL1 | Plasmid harboring PCDR1-lacZ fusion for ACT1 locus integration | Gaur et al. (11) |
| pCPL51 | Plasmid harboring PCDR1-lacZ fusion for CDR1 locus integration | This study |
| pCPL52 | Plasmid harboring CDR1-lacZ fusion for CDR1 locus integration | This study |
| pCPL62 | Plasmid harboring in frame CDR1-lacZ fusion for CDR1 locus integration | This study |
| Oligonucleotides | ||
| CDR1F | Forward primer for CDR1 promoter amplification | 5′-GATCGGGCCCTCGTTACTCAATAAGTAT-3′ |
| CDR1R | Reverse primer for CDR1 promoter amplification | 5′-AGCTCTCGAGTTCTTTTTGACCTTTTAAAG-3′ |
| CDR1-F-RML | Forward primer for CDR1 promoter amplification | 5′-GCTTCGCCTCAACTTCTTATAAAGTTTTGAAAG-3′ |
| CDR1-R-RML | Reverse primer for CDR1 promoter amplification | 5′-CGTGGTATTCATTAATGGAATGGTTTTCTGAAG-3′ |
| CDR29 | Forward primer for amplification of CDR1 downstream region | 5′-AATTCTGCAGTTTGTTTTTTGACATGGTGGTATC-3′ |
| CDR30 | Reverse primer for amplification of CDR1 downstream region | 5′-TCGTGCCGCGGAACACTTTTTCTTTCTAATTATAA-3′ |
| CDR32 | Forward primer for amplification of CDR1 C-terminal region | 5′-ATTTGGTACCATACATTAAATTTGCTGGTGGG-3′ |
| CDR31 | Reverse primer for amplification of CDR1 C-terminal region | 5′-GTTTGGATCCTTTCTTATTTTTTTTCTCTCTGTTACCC-3′ |
| CT-CDR1-R-RML | Reverse primer for amplification of CDR1 C-terminal region | 5′-GTTTCTCGAGTTTCTTATTTTTTTTCTCTCTGTTACCC-3′ |
| A-del pCPL52-F | Forward primer for in-frame translational fusion of CDR1-ORF with lacZ | 5′-GAGAGAAAAAAAATAAGAAACTCGAGATGAACATGACTGAAAAAATTCAAAC-3′ |
| A-del pCPL52-R | Reverse primer for in-frame translational fusion of CDR1-ORF with lacZ | 5′-GTTTGAATTTTTTCAGTCATGTTCATCTCGAGTTTCTTATTTTTTTTCTCTC-3′ |
| KM1 | Forward primer for amplification of CDR1 N-terminal region | 5′-CTTTTCCACTGGTAACTACT-3′ |
| KM2 | Reverse primer for amplification of CDR1 N-terminal region | 5′-CAATAAACCTGCTGACGAG-3′ |
a
Restriction sites introduced into the primers are underlined, while the flanking bases are written in italics.