Table 1 Primers pairs for introduction of selected mutations in pET-25b(+) expression vector

Pair #1

Reverse Eco72 I

5^′ GAGCAGCAGACCAGCAGCAGCGGTCGGCAGCAC GTG TTC CATATGTATATCTCCTT 3^′

Forward Eco72 I

5^′ CGCTGCTGCTGGTCTGCTGCTCCTCGCTGCCCAGCCGGC 3^′

Pair #2

Reverse Eco147 I

5^′ GGGGTCTTCCGGGGCGAGTTCTGGCTGGCTAGG CCT TTTGATCTCGAGTGCGG 3^′

Forward Eco147 I

5^′ AGAACTCGCCCCGGAAGACCCCGAGGATGTCGAGCACCA 3^′