Nickel quercetinase, a “promiscuous” metalloenzyme: metal incorporation and metal ligand substitution studies

BMC Biochemistry

Table 3 Oligonucleotides used for gene amplification and site-directed mutagenesis

Name	Nucleotide sequence (5′ → 3′)	Application
quedfor	TGACCATCGAATACGCCACCCGTCACC	Amplification of queD
quedrev	GACGTCGTACTGCTCGGGCACGGTC	Amplification of queD
strepfor	ATAAGAATGCGGCCGCATTAGAATGGAGCCACCCGCAGTTCGAAAAATGAGATCCGGCTGCTAACAAAGCCCGAAAGG	Exchange of His₆-tag for StrepII-tag
streprev	ATAAGAATGCGGCCGCAAGCTTCCTTCCCTCGATACTCCCGGTGTGC	Exchange of His₆-tag for StrepII-tag
gfpfor	TATAAGCTTGCCGGTGCGGAAAATC	Amplification of the EGFP gene
gfprev	TATAGCGGCCGCCTGGTACAGTTCATCCATG	Amplification of the EGFP gene
H69Afor	CCTCGCACGCGGACACCTACG	Exchange of His69 for Ala
H69Arev	CGGCGGGGATCACCTCGCCCTTG	Exchange of His69 for Ala
H71Afor	CGCCCACTCGGCCGCGGACACCTACGAGGTCTTC	Exchange of His71 for Ala
H71Arev	GGGATCACCTCGCCCTTGGGGCCCTCGCAG	Exchange of His71 for Ala
H115Afor	ATGGAACGCCACCACTCGCAGGTCG	Exchange of His115 for Ala
H115Arev	GCGGTAGGCGGCCACGCAGTTCTTCGGTACGAAGC	Exchange of His115 for Ala
E76Hfor/E76Dfor	CGTCTTCTACATCACCCAGGGC	Forward primer for the exchange of Glu76 for His or Asp
E76Hrev	TGGTAGGTGTCCGCGTGCGAGTGG	Reverse primer for the exchange of Glu76 for His
E76Drev	TCGTAGGTGTCCGCGTGCGAG	Reverse primer for the exchange of Glu76 for Asp

ISSN: 1471-2091