Table 2 Polymerase chain reaction primers used in this study
From: Aspergillus oryzae-based cell factory for direct kojic acid production from cellulose
Primers | Sequence (5′-3′) | Template DNA |
|---|---|---|
ascI-kojR-F | AAAAAGGCGCGCCATGTCGTTGAATACCGACGATTCC | A. oryzae genome DNA |
notI-FLAG-kojR-R | AAAGCGGCCGCTTACTTGTCATCATCGTCCTTATAGTCTCTATATCTCTGACCACCTGC | |
CBHI-F | CCAAACCACCCAAAGGGCGCGCCATGTATCGGAAGTTGGCCGTCATC | synthesized gene |
CBHI-R | GAAAGTACATGTCGAGCGGCCGCTTACAGGCACTGAGAGTAGTAAG | |
psvaA-ascI-CBHI-F | GGCGCGCCAAAAAATGTATCGGAAGTTGGCCGTCA | pISI-CBHI |
tsvaA-notI-CBHI-R | AAATGATGCGGCCGCTTACAGGCACTGAGAGTAGT | |
pIS1-psvaA-F | CACAACACTCTCGACCTGCAGCGGTTTACACCGAAGACCGG | A. oryzae genome DNA |
CBHI-ascI-psvaA-R | ATTTTTTGGCGCGCCCTTGCGAGCAGGGGGATAAT | |
CBHI-notI-tsvaA-F | GCGGCCGCATCATTTTCCCGCTTTGATCTGGTCGGTTCCC | A. oryzae genome DNA |
pIS1-tsvaA-R | GGTACCCGGGGATCCTCTAGATCAAGCATAACTACAACAGGGCAAGGAATACGG | |
EGI-F | CCAAACCACCCAAAGGGCGCGCCATGGCGCCCTCAGTTACACTGC | T. reesei cDNA |
EGI-R | GAAAGTACATGTCGAGCGGCCGCCTAAAGGCATTGCGAGTAGTCTG | |
phlyA-ascI-EGI-F | GGCGCGCCAAAAAATGGCGCCCTCAGTTACACTGC | pISI-EG |
thlyA-notI-EGI-R | AGAGAGAGCGGCCGCCTAAAGGCATTGCGAGTAGT | |
pIS1-phlyA-F | CACAACACTCTCGACCTGCAGTACAGCATGGTCTGGATTCCAATCCACGCAGC | A. oryzae genome DNA |
EGI-ascI-phlyA-R | ATTTTTTGGCGCGCCGGTGTTGTGGTGTGAAGGGTGATTGATGTGAGACC | |
EGI-notI-thlyA-F | GCGGCCGCTCTCTCTCCCCTATACGTGATACCGTA | A. oryzae genome DNA |
pIS1-thlyA-R | GGTACCCGGGGATCCTCTAGAGATGCAAATTGGAGTTAAAT | |
sCutL-F | CCAAACCACCCAAAGGGCGCGCCATGCATCTTGCTATCAAGTCTC | A. oryzae genome DNA |
sCutL-R | TCACTAGTTCTCTCAACCAGAGCATTGCTGGG | |
N28-F | TTGAGAGAACTAGTGATGACAACTTGGTTGGTGGCATG | pISI-GFP (Adachi et al.[14]) |
N28-R | AGTTCATCCTCGACACGCTTGGCGCTGTTGG | |
BGL-F | GTGTCGAGGATGAACTGGCGTTCTCTCCTC | pdU-PGAGBGL (Yamada et al.[15]) |
BGL-R | GAAAGTACATGTCGAGCGGCCGCTTACTTGTCATCGTCATCCTTG | |
pIS1-svaA-CBHI-F | GGCTTTCCCCGTCAAGCTCT | pISI-svaA-CBHI |
pIS1-svaA-CBHI-R | GGCGAACGTGGCGAGAAAGG | |
phlyA-into-pIS1-svaA-CBHI-F | CTCGCCACGTTCGCCTACAGCATGGTCTGGATTCCAATCCACGCA | pISI-hlyA-EGI |
connect-thlyA-and-psodM-R | TATTTAAATATCGATGATGCAAATTGGAGTTAAATATTAACTAAC | |
connect-psodM-and-thlyA-F | ATCGATATTTAAATATTATGTACTCCGTACTCGGTTGATTATTAA | pIS1-BGL1 |
tglaB-into-pIS1-svaA-CBHI-R | TTGACGGGGAAAGCCGGATGTAGTATGTATACTTAGTTTGATTGC | |
hlyA-EGI-svaA-CBHI-sodM-BGLI-F | TTTATATCCAAGATCACCTGCAGCGGTTTACACCGAAGACCGGTA | pISI-CBHI-EGI-BGL1 |
hlyA-EGI-svaA-CBHI-sodM-BGLI-R | TGCCAAGAGAAGCTTGGCGTAATCATGGTCATAGCTGTTT | |
sC-F | AAGCTTCTCTTGGCAATAGCTGCCC | pUSC (Yamada et al.[16]) |
sC-R | GATCTTGGATATAAAAATCCAAATATGGCTCCTCG | |
kojR-RT-PCR-F | TGGTGCAATCAGCGAAGGA | wild type and engineered A. oryzae cDNA |
kojR-RT-PCR-R | AGACTACTCTCCTGCATCATGCC | |
kojA-RT-PCR-F | ATCCGAAGGCGAATGGTTT | wild type and engineered A. oryzae cDNA |
kojA-RT-PCR-R | ATGAACCCAGCGTCGCTATT | |
kojT-RT-PCR-F | CATGGTGCCGCATATTTACTTC | wild type and engineered A. oryzae cDNA |
kojT-RT-PCR-R | AATGGACACAATGGGTTGCC | |
ß-Tubulin-RT-PCR-F | GCCAGTGTGGTAACCAAATAGGT | wild type and engineered A. oryzae cDNA |
ß-Tubulin-RT-PCR-R | TAAACACCGGAGCCGTCAA |