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Table 1 Oligonucleotides used for cloning and mutagenesis

From: Aglycone specificity of Thermotoga neapolitana β-glucosidase 1A modified by mutagenesis, leading to increased catalytic efficiency in quercetin-3-glucoside hydrolysis

Primer

Direction

Mutation

Sequencea, b

1

Forward

N/A

5'-TATTCTTATCATATG AAAAAGTTTCCCGAAGGGTTC

2

Reverse

N/A

5'-TATTCTTATCTCGAG ATCTGTTAGTCCGTTGTTTTTG

3

Forward

N/A

5'-AATACGACTCACTATAGG

4

Reverse

N/A

5'-CTAGTTATTGCTCAGCGG

5

Forward

F219L

5'GACGGAAAGATAGGGATTGTTTTA AACAACGGATACTTCGA

6

Forward

N221S

5'-GGGATTGTTTTCAACAGC GGATACTTCGAACCTGC

7

Forward

G222M

5'-GATTGTTTTCAACAACATG TACTTCGAACCTGCAAG

8

Forward

G222Q

5'- GGATTGTTTTCAACAACCAA TACTTCGAACCTGCAAGTGAGAGAG

9

Reverse

F219L

5'- GACGGAAAGATAGGGATTGTTTTA AACAACGGATACTTCGA

10

Reverse

N221S

5'- GCAGGTTCGAAGTATCCGCT GTTGAAAACAATCCC

11

Reverse

G222M

5'-CTCTCTCACTTGCAGTAC CGAAGTATTGGTTGTTGAA

12

Reverse

G222Q

5'- CTCTCTCACTTGCAGGTT CGAAGTATTGGTTGTTGAAAACAATCC

  1. aThe restriction sites for cloning are underlined. bInduced changes are shown in bold.