Table 2 Primers used in this study
Primers | Sequence (5′ to 3′) | Restriction site/utilisation |
|---|---|---|
LYS5-P1 | ATAAGAATGCGGCCGCCGACTAAATTTCGACCCAC | NotI, LYS5 disruption |
LYS5-P2 | CGATTACCCTGTTATCCCTAGCGTAACTCGCTACTAGGCCGCCACC | I-SceI, LYS5 disruption |
LYS5-T1 | CGTAGGGATAACAGGGTAATATAGCGTAACTATAACGGTCCTAAGGTAGCGAAGGCGTTGGTGCTCTCTCGGAAGTAG | I-SceI, LYS5 disruption |
LYS5-T2 | ATAGTTTAGCGGCCGCAAAAATGTCCGCCATTGAGTGTTG | NotI, LYS5 disruption |
LPR-R | GCTAGATAGAGTCGAGAATTACCCTG | LYS5 prototrophy |
LYS5PR | TCGGTGCGTGTGAAAGACAC | LYS5 prototrophy |
preTEYK Fw | GTGTTTGACATTTTGTTTTGTGTGAGT | Verification of EYK1 disruption |
postPEYK Rv | TACACACTCACACTCACCAGAACATC | Verification of EYK1 disruption |
ClaI-pEYD1-Fw | CCCATCGATGGAAACCTTAATAGGAGACTACTTCC | pEYD1 cloning |
no AvrII-pEYD1-Fw | CCTCGTGTCCGGGCTAGGGCAGAAACAGCTC | pEYD1 cloning, pEYD1 verification |
no AvrII-pEYD1-Rev | GAGCTGTTTCTGCCCTAGCCCGGACACGAGG | pEYD1 cloning |
BamHI-pEYD1-Rev | TGTGTATGTGTGTGTGTGTGTGTGTGTGTGTTTG | pEYD1 cloning |
pTEF-internal-Fw | TCTGGAATCTACGCTTGTTCA | pTEF verification |
EYK300-Fw | GCATCTACTTTTCTCTATACTGTACGTTTCAATCTGGG | pEYK1-3AB, pHU8EYK verification |
CalB-prepro-Fw | ATGAAGCTGCTGTCTCTGACC | CalB verification |
CalB-internal-Rev1 | CCACCTTAGATCGAATAGAAGGG | CalB verification |
CalB-Rev | TTAAGGGGTGACAATACCAGAAC | CalB verification |
ACT-F | TCCAGGCCGTCCTCTCCC | qPCR |
ACT-R | GGCCAGCCATATCGAGTCGCA | qPCR |
CalB-internal-Fw | TCTCTGCTCCTTCTGTGTGG | qPCR |
CalB-internal-Rev2 | GTCGAACAGAGGTCCACAGA | qPCR |