Table 2. Plasmids used in the present study.
UTR, untranslated region.
| Name | Description | Ref/origin |
|---|---|---|
| pGBDU-C1 and C3 | Two-hybrid plasmid, Ura-marked | [34] |
| pGAD-C1 | Two-hybrid plasmid, Leu-marked | [34] |
| pKA142 | pGEX4T1 | Amersham |
| pKA260 | Nucleotides 1460–2192 of Sla1 corresponding to region HD1+HD2 were amplified by PCR. The product was then cloned into pGBDU-C3. | [14] |
| pKA263 | Nucleotides 522–1065 of Lsb5, including the GAT domain to end of gene in pGAD-C1 | [14] |
| pKA267 | pGEX2T+full-length Lsb5 amplified by PCR with flanking BamHI and BglII restriction sites | [14] |
| pKA273 | pQE31 His-tagging vector (Qiagen) with nucleotide region 1460-2192 of Sla1, which was subcloned using restriction sites BamHI and PstI from pKA260 | This study |
| pKA312 | PGBDU-C1+LSB5(1–1062) | This study |
| pKA316 | PEGFP-C2+LSB5 | This study |
| pKA325 | PGAD-C1+LAS17(292–536) isolated from screen | |
| pKA342 | pKA260 with stop codon introduced at nucleotide 1733 of Sla1 using oligonucleotides oKA329 and 330 | This study |
| pKA343 | pKA260 with stop codon at nucleotide 1927 using oligonucleotides oKA331 and 332 | This study |
| pKA346 | pGAD-C1+Lsb5(301–354). Lsb5 from nucleotide 901 was cloned after PCR amplification from genomic DNA using oligonucleotides oKA346 and 347 | This study |
| pKA365 | pGAD-Las17(292–422). Stop codon in pKA325 introduced using oKA323 and 324. | This study |
| pKA366 | pGAD-Las17(292–384). Stop codon in pKA325 introduced using oKA325 and 326. | This study |
| pKA367 | pQE31 His-tagging vector with nucleotides 1460–2192 of Sla1, subcloned using restriction sites BamHI and PstI from pKA342. | This study |
| pKA369 | pGEX6P2+Arf3 lacking the N-terminal 14 amino acids | S. Munro |
| pKA370 | pGEX6P2+Arf3 lacking the N-terminal 14 amino acids and GDP-locked (T31N) | S. Munro |
| pKA371 | pGEX6P2+Arf3 lacking the N-terminal 14 amino acids and GTP-locked (Q71L) | S. Munro |
| pKA372 | pRS316+GFPArf3 | S. Munro |
| pKA384 | pNB701. pRS316 plasmid modified for GFP tagging under CUP1 control. | N. Bryant |
| pKA409 | pKA263 with stop codon introduced at nucleotide 1036 of Lsb5 using oligonucleotides oKA443 and 444 | This study |
| pKA410 | pKA346 with stop codon introduced at nucleotide 1036 of Lsb5 using oligonucleotides oKA443 and 444 | This study |
| pKA416 | pKA384+Lsb5. Lsb5 PCR amplified using oligonucleotides oKA441 and 442 with 3′-CUP1 and 5′-UTR flanking from a plasmid containing full-length Lsb5. | This study |