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Table 5 Primers and locus tags of the nitrilase genes in the predicted mandelonitrile hydrolase subgroup used in this study

From: Discovery and characterization of a highly efficient enantioselective mandelonitrile hydrolase from Burkholderia cenocepacia J2315 by phylogeny-based enzymatic substrate specificity prediction

Sequence Accession no.

Locus_tag

Source

Primer pair (sequence 5’→ 3’)

Restriction enzymes

CP000699

Swit_0987

Sphingomonas wittichii RW1a

Forward: CATATGTCGGACGGTCCGTTCAAGGTTG

NdeI/HindIII

Reverse: AAGCTTTTAGGAACCGGCCTTTTCGGCGA

CU234118

BRADO4539

Bradyrhizobium sp. ORS 278a

Forward: GGGAATTCCATATGGGACTGGCACATCCG

NdeI/HindIII

Reverse: GAAAAAAGCTTACCCGCCAGCCGCGACCT

CP000494

BBta_4766

Bradyrhizobium sp. BTAi1b

Forward: GGATCCATGGGACTGGCACATCCGAAATAC

BamHI/HindIII

Reverse: AAGCTTCAGATGGATTGATCGGGCCGCGCG

AM747720

BCAL2585

Burkholderia cenocepacia J2315b

Forward: GGATCCATGACCATCAATCACCC

BamHI/HindIII

Reverse: AAGCTTAAGCGGGTGTGACGC

CP000555

Mpe_A0993

Methylibium petroleiphilum PM1a

Forward: CATATGCCGGTTTCGCACCCCAAG

NdeI/HindIII

Reverse: AAAAAAGCTTAAGCCGTGCGGCGCGCGGTC

BAD58116

nfa32690

Nocardia farcinica IFM 10152a

Forward: CATATGAGTCAGCGAGACAGTTTCCG

NdeI/HindIII

Reverse: ATATAAGCTTCCGCACCGCGGGTTCGGCGT

NZ_AAUW01000019

SIAM614_26356

Stappia aggregata IAM 12614a

Forward: CATATGAAAGCTATCAAGGTTGCCGCCGTTC

NdeI/BamHI

Reverse: GGATCCCTACTCCTCGACCTCAAAAGGCCGT

  1. a The gene was cloned into a pET28a(+) expression vector using E. coli BL21 (DE3) as host strain.
  2. b The gene was cloned into a pQE30 expression vector using E. coli M15 as host strain.
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BMC Biotechnology

ISSN: 1472-6750

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