Table 2.
Oligonucleotides used for PCR amplification in this study
| Primer name | Sequence (5′-3′) |
|---|---|
| Primers for DNA fragment PCR to obtain a partial sequence of genomic DNA of the acid proteinase | |
| 12 ND-F |
AACGATATCGAGTACTATGGT |
| M.cir-2R |
TTAAAGACTTCATAGTTGTTCTT |
| Primer for 3′-RACE PCR (Gene-specific primer) | |
| GSP-Mucor-1 F |
GATGGTCGTGCCTGGTCTATCCAAT |
| Primer for 5′-RACE PCR (gene-specific primer) | |
| GSP-Mucor-2R |
CATTGTCTCTGGCACCGTATTGAGCAGC |
| Primers for full-length cDNA and recombinant plasmids | |
| APMC-EcoNaeI-F |
ATGGAATTCGCCGGCGCTACTACTGATGCCACTGGTACTGTCCCCG |
| APMC-F |
AGGAATTCTTCTCATTAGTCTCTTCTTG |
| APMC-Met-F |
ATGGAATTCATGAAATTCTCATTAGTCTCTTCTTGTGTC |
| MCAP-3 F |
TATCTCGAGaaaagaGCTCCCAGTGGTAGCAAGAA |
| XhoI-N-MCAP-F |
TATCTCGAGaaaagaATGAAATTCTCATTAGTCTCTTCTTGTG |
| APMC-NotI-R |
AAAGCGGCCGCGACAGATTTGGCAATTT |
| APMC-Stop-R |
GTGATTTATAGATAGATAGATGAAATGTACCAAA |
| Primers to identify clones containing recombinant plasmids | |
| pGAP-F |
GTCCCTATTTCAATCAATTGAACAAC |
| AOX1pGAP-Rev | CAAATGGCATTCTGACATCCTC |
The underlined sequences (GAATTC; EcoRI, CTCGAG; XhoI and GCGGCCGC; NotI) represent the additional restriction sites at the 5′ ends of forward and reverse primers. The lowercase letters indicate the Kex2 cleavage sites. The primers (for First-strand cDNA synthesis, 3′-RACE cDNA and 5′-RACE cDNA) provided in the SMART RACE cDNA Amplification Kit (Clontech) are not described in the table.