Efficient Optimization of Gluconobacter oxydans Based on Protein Scaffold-Trimeric CutA to Enhance the Chemical Structure Stability of Enzymes for the Direct Production of 2-Keto-L-gulonic Acid
Table 2
Primers used in this study.
Primer
Sequences of primers (5′-3′)
Restriction enzyme
tufB-F1
CGAGCTCGTACGATGGTAAGAAATCCACTG
SacI
tufB-R1
CGGGGTACCCGTCTTTCTCCAAAACCCC
KpnI
SH3-F
CGGGGTACCGCCGAGTATGTGCGCGCCCT
KpnI
SH3-R
GGGTCGTGAGCTTCATGTACTTCTCCACGTACGGCACCG
—
sdh-GGGGS-sndh-F
GTACGTGGAGAAGTACATGAAGCTCACGACCCTGCTGC
—
sdh-GGGGS-sndh-R
CGCGGATCCTCACGCCGCGGAAATCCGC
BamHI
tufB-F2
CGCGGATCCGTACGATGGTAAGAAATCCACTG
BamHI
tufB-R2
GCAGGGCCGGCGGCGGCGTCTTTCTCCAAAACCCCGCT
—
SH3lig-(GGGGS)2-cutA-F
AGCGGGGTTTTGGAGAAAGACGCCGCCGCCGGCCCTGC
—
SH3lig-(GGGGS)2-cutA-R
CTAGTCTAGATCACTTCTTCGTCTCCTCGATCAGC
XbaI
tufB-F3
CTAGTCTAGATGCAGATCCGGTGGCCATGTTC
XbaI
tufB-R3
TTCGGCGTGTTCCAAGCCATCGTCTTTCTCCAAAACCCCGCT
—
pqqABCDE-F
CGGGGTTTTGGAGAAAGACGATGGCTTGGAACACGCCG
—
pqqABCDE-R
AAAACTGCAGTTACATTCTTCGGTAAACAAAGT
PstI
Restriction sites used for cloning are in bold and are underlined.
