Table 1 Primers using in this study
Description | Primer name | Primer sequence (5′–3′) | Restriction site |
|---|---|---|---|
Preparation for expression vectors | |||
 pPIC9-GFP(RSIR) | RSIR F | CCGCTCGAGAGATCTATCAGAGTGAGCAAGGGCGAG | XhoI |
RSIR R | CCGGAATTCTTACTTGTACAGCTCGTC | EcoRI | |
 pPIC9-GFP(RSII) | RSII F | CTCTCGAGAGATCTATAATAGTGAGCAAGGG |  |
RSII R | CCCTTGCTCACTATTATAGATCTCTCGAGAG | Â | |
 pPIC9-GFP(ISIR) | ISIR F | CTCTCGAGATATCTATAAGAGTGAGCAAGGG |  |
ISIR R | CCCTTGCTCACTCTTATAGATATCTCGAGAG | Â | |
 pPIC9-GFP(RSRR) | RSRR F | CTCTCGAGAGATCTAGAAGAGTGAGCAAGGG |  |
RSRR R | CCCTTGCTCACTCTTCTAGATCTCTCGAGAG | Â | |
 pPIC9-GFP(RSRI) | RSRI F | CTCTCGAGAGATCTAGAATAGTGAGCAAGGG |  |
RSRI R | CCCTTGCTCACTATTCTAGATCTCTCGAGAG | Â | |
 pPIC9-GFP(ISRR) | ISRR F | CTCTCGAGATATCTAGAAGAGTGAGCAAGGG |  |
ISRR F | CCCTTGCTCACTCTTCTAGATATCTCGAGAG | Â | |
 pPIC9-GFP(RSKR) | RSKR F | CTCTCGAGAGATCTAAGAGAGTGAGCAAGGG |  |
RSKR R | CCCTTGCTCACTCTCTTAGATCTCTCGAGAG | Â | |
 pPIC9-GFP(RSKI) | RSKI F | CTCTCGAGAGATCTAAGATAGTGAGCAAGGG |  |
RSKI R | CCCTTGCTCACTCTCTTAGATATCTCGAGAG | Â | |
 pPIC9-GFP(ISKR) | ISKR F | CTCTCGAGATATCTAAGAGAGTGAGCAAGGG |  |
ISKR R | CCCTTGCTCACTATCTTAGATCTCTCGAGAG | Â | |
 pGAPZ B-Kex2 | Kex2 F | CCGGAATTCCGGATGTATTTGCCAGCACTT | EcoRI |
Kex2 R | CCGCTCGAGCGGTTACAATGCCGCACGTTT | XhoI | |
 pGAPZ B-furin | Furin F | CCGGAATTCCGGATGGAGCTGAGGCCCTGG | EcoRI |
Furin R | CCGCTCGAGTTAGAGGGCGCTCTGGTCTT | XhoI | |
Preparation for disruption vectors | |||
 pYPS1Δ | Left arm F | TGCACTGCAGTAGCCGTTCCCGCGTGAAGA | PstI |
Left arm R | CCGCTCGAGCGGACTAGCATATGTGGATTCTAG | XhoI | |
Right arm F | GGAAGATCTTCCTCCATCCTCTTTGGAGGTGTG | BglII | |
Right arm R | TGCACTGCAGTGCACTATACTATACACACG | PstI | |
 pKex2Δ | Left arm F | TGCACTGCAGTGCAATGTATTTGCCAGCAC | PstI |
Left arm R | CCGCTCGAGCGGTGGTGGATGAAGCCCTTTAAT | XhoII | |
Right arm F | GGAAGATCTTCCACGGATATGGCAAGATCGATG | BglII | |
Right arm R | TGCACTGCAGTGCATTACAATGCCGCACGTTTGGG | PastI | |
Identification for disrupted strains | |||
 GS115-ΔYps 1 | YI F | CAGTATGACAATTTGCCAGC |  |
YI R | TCCCCCGGTGTAATATGTTG | Â | |
YO F | GGACTTCAGCGTTCTGAGGG | Â | |
AOX1 R | GCAAATGGCATTCTGACATCC | Â | |
 GS115-ΔKex 2 | KI F | GGCTTGCTCTGCTGTGATGA |  |
KI R | AGATCTTTGGCCTCCTGGG | Â | |
KO F | CACCTGAAACTTAATACTCT | Â | |
AOX1 R | GCAAATGGCATTCTGACATCC | Â | |
Identification for GS115-furin and GS115-Kex2 strains | |||
 GS115-furin | AOX1 F | GACTGGTTCCAATTGACAAGC |  |
AOX1 R | GCAAATGGCATTCTGACATCC | Â | |
 GS115-Kex2 | GAP F | GTCCCTATTTCAATCAATTGAA |  |
Kex2 R | GCCTCCTGGGTCAATTCATA | Â | |
RT-PCR | Â | ||
 β-actin | β-actin F | CTCCAATGAACCCAAAGTCCAAC |  |
β-actin R | GACAAAACGGCCTGAATAGAAAC |  | |
 Furin | RT-furin F | GGCATTGTGGTCTCCATTCT |  |
RT-furin R | GCAGTTGCAGCTGTCATGTT | Â | |
 Kex2 | RT-Kex2 F | GGCATTGTGGTCTCCATTCT |  |
RT-Kex2 R | GCAGTTGCAGCTGTCATGTT | Â | |