Table 1 Primers for gene cloning and expression.
From: Clonostachys rosea demethiolase STR3 controls the conversion of methionine into methanethiol
Genes | Primers | Sequence |
|---|---|---|
Homology-based cloning | ||
AAT1 | AAT1-5F | 5′-ATHAAYYTNGGNGTNGGNGCNTA-3′ |
AAT1-3R | 5′-NARNCCNGTRTANGCRAACAT-3′ | |
AAT1-3R-22 | 5′-CCCATRTTYTTNGCRAANSWYTG-3′ | |
AAT2 | AAT2-5F | 5′-YTNGGNATHGGNGCNTAGMGNGA-3′ |
AAT2-3R | 5′-DATNCKNCCNSWCATHGTNC-3′ | |
AAT2-5F-22 | 5′-CAYGCNTGYGCNCAYAAYCCNAC-3′ | |
AAT2-3R-22 | 5′-GGNGGRTTNSWDATYTCNSWNC-3′ | |
ARO8 | ARO8-5F2 | 5′-ATHWSNYTNGGNGGNGGNYTNCC-3′ |
ARO8-3R-23 | 5′-RAACCANSWNCCNCKNGCNAC-3′ | |
ARO8-4R | 5′-RTARTANGGYTCRTCYTCDATDAT-3′ | |
BAT | BAT-5F1 | 5′-TAYGCNACNGARTGYTTYGARGG-3′ |
BAT-3R1 | 5′-RTCNARNGGNGCNGTDATNA-3′ | |
BAT-5F2 | 5′-GNAARYTNMGNYTNTTYMGNCC-3′ | |
BAT-3R2 | 5′-RTARTTNGCNCCNACYTT-3′ | |
STR3 | STR3-5F | 5′-MGNWSNGGNAAYCCNACNMG |
STR3-3R | 5′-ARNSWRTTNACRCANCCRAA-3′ | |
STR3-3R2 | 5′-NCCNGTYTCRAANSWNARNACNGC-3′ | |
Full-length cDNA cloning | ||
AAT1 | AAT1-3RA1 | 5′-CCCTCGACAGGCTCGCCATTACCCAG-3′ |
AAT1-3NRA2 | 5′-GTGCTGAGTTCCTTCAGCGCTGGTACAG-3′ | |
AAT1-5RA1 | 5′-CACTGGTCCTGGCTGGGGTCAACA-3′ | |
AAT1-5NRA2 | 5′-GAGGAGTGTTCTTGAGGTCCTCAACCAG-3′ | |
AAT2 | AAT2-3RA1 | 5′-GAGCTGCTCATCGCCCAGAGCTTC-3′ |
AAT2-3NRA2 | 5′-GTGCTGGCTGCTTCCACGCCATCACCTC-3′ | |
AAT2-5RA1 | 5′-CTCACTCATTAGGCACCCCAGGC-3′ | |
AAT2-5NRA2 | 5′-CACAGGAAACAGCTATGACCATGATTAC-3′ | |
ARO8 | ARO8-3RA1 | 5′-TGACCTCTCCATCGCCCTCAACTAC-3′ |
ARO8-3NRA2 | 5′-GAACACACCGAGCTGGTCTGCCGCCCTC-3′ | |
ARO8-5RA1 | 5′-TCGTCCATTGCCTCGGGGATGAGTC-3′ | |
ARO8-5NRA2 | 5′-GAACATGTTGATTCCAAGAGGGGCGAATG-3′ | |
ARO8-2 | ARO8-23RA1 | 5′-AGCTCCATGCAGCAGCGAGACCTTC-3′ |
ARO8-23NRA2 | 5′-CCAGCCGGACTGGCTCATCCCTCAAGGGTG-3′ | |
ARO8-25RA1 | 5′-AGCGCAGAAGCTGGGCTGAGCCAC-3′ | |
ARO8-25NRA2 | 5′-CACTCTGGCCGTCTCGGAGATCGTGCTTG-3′ | |
BAT | BAT-3RA1 | 5′-GAGCAGCGCGGATACTCCCTCTAC-3′ |
BAT-3NRA2 | 5′-CCAGGCTCAGCCCTCCTCTATGTTATTG-3′ | |
BAT-5RA1 | 5′-GTGACGTACTGCTCCTCGCCGAAG-3′ | |
BAT-5NRA2 | 5′-CAGGTTCTGCTGGTGGCCCCTGGAGCTG-3′ | |
STR3 | STR3-3RA1 | 5′-TCCTCGAAACACCTACAAACCCTC-3′ |
STR3-3NRA2 | 5′-CCAACAATCGCCCGGAATGCCCACGAAG-3′ | |
STR3-5RA1 | 5′-GAATGGCAAGAGTCTTGACTCCTC-3′ | |
STR3-5NRA2 | 5′-TCATTGGGTGACAGACCGCAGCCAGTTG-3′ | |
STR3 expression in S. cerevisiae | ||
STR3 | YSTR3-5H | 5′-CCCAAGCTTAACACAATGTCTGCCCCGCCTCCGCCAAATG-3′ |
YSTR3-3BH | 5′-CGCGGATCCTTAGTGGTGGTGGTGGTGGTGTTTGGCTGTGCGTGGAGTTCGTC-3′ | |
F5 | 5′-AGTATCAACAAAAAATTGTTA-3′ | |
R3 | 5′-GAATGTAAGCGTGACATAACTAA-3′ | |
