Enhancement of Polymerase Activity of the Large Fragment in DNA Polymerase I from Geobacillus stearothermophilus by Site-Directed Mutagenesis at the Active Site

Table 1

Oligonucleotides used in substitution mutagenesis for 10 residues of Bst DNA pol LF.

Mutagenesis	Primer name	Sequence
G310A	F (5′-3′)	GTGGTGCACCCCGTGACGGCGAAAGTGCACACGATGTTCAATCAG
	R (5′-3′)	CTGATTGAACATCGTGTGCACTTTCGCCGTCACGGGGTGCACCAC
G310L	F (5′-3′)	GTGGTGCACCCCGTGACGCTCAAAGTGCACACGATGTTCAATCAG
	R (5′-3′)	CTGATTGAACATCGTGTGCACTTTGAGCGTCACGGGGTGCACCAC
D540A	F (5′-3′)	CGCCTGTTGCTGCAAGTGCATGCAGAACTGATTTTGGAGGCGCCGAAAGA
	R (5′-3′)	CTCTTTCGGCGCCTCCAAAATCAGTTCTGCATGCACTTGCAGCAACAGGCG
D540E	F (5′-3′)	CGCCTGTTGCTGCAAGTGCATGAAGAACTGATTTTGGAGGCGCCGAAAGA
	R (5′-3′)	CTCTTTCGGCGCCTCCAAAATCAGTTCTTCATGCACTTGCAGCAACAGGCG
R412A	F (5′-3′)	CGAAGAAGACGTGACAGCCAACATGCACGCCAAGCGAAGGCCGTCAATT
	R (5′-3′)	AATTGACGGCCTTCGCTTGGCGTGCCATGTTGGCTGTCACGTCTTCTTCG
R412E	F (5′-3′)	CGAAGAAGACGTGACAGCCAACATGGAACGCCAAGCGAAGGCCGTCAATT
	R (5′-3′)	AATTGACGGCCTTCGCTTGGCGTTCCATGTTGGCTGTCACGTCTTCTTCG
K416A	F (5′-3′)	GTGACAGCCAACATGCGCCGCCAAGCGGCAGCCGTCAATTTTGGCA
	R (5′-3′)	TGCCAAAATTGACGGCTGCCGCTTGGCGGCGCATGTTGGCTGTCAC
K416D	F (5′-3′)	GTGACAGCCAACATGCGCCGCCAAGCGGATGCCGTCAATTTTGGCA
	R (5′-3′)	TGCCAAAATTGACGGCATCCGCTTGGCGGCGCATGTTGGCTGTCAC
G310A-D540E	F (5′-3′)	GTGGTGCACCCCGTGACGGCGAAAGTGCACACGATGTTCAATCAG
	R (5′-3′)	CTGATTGAACATCGTGTGCACTTTCGCCGTCACGGGGTGCACCAC
G310L-D540E	F (5′-3′)	GTGGTGCACCCCGTGACGCTCAAAGTGCACACGATGTTCAATCAG
	R (5′-3′)	CTGATTGAACATCGTGTGCACTTTGAGCGTCACGGGGTGCACCAC

Underlined sequences indicate the mutant codons.