TABLE 1.

Oligonucleotides used in this studya

PrimerSequence (5′ → 3′)
VIM-2-EXP/fwdGGAATTCCATATGTTCAAACTTTTGAGT
VIM-2-EXP/revGGCTCGAGGATCCTGCTACTCAACGACTG
VIM-random 61(+)CAACGCAGTCGNNNGATGGCCCAG
VIM-random 61(−)CTGCGCCATCNNNCGACTGCCTTG
VIM-random 64(+)GTTTGATGGCNNNGTCTACCCG
VIM-random 64(−)CGGCTAGACNNNGCCATCAAAC
VIM-random 67(+)GGCGCAGTCNNNCCGTCCAATG
VIM-random 67(−)CATTGGACGGNNNGACTGCGCC
VIM-random 87(+)GATTGATACAGCGNNNGCTGCGAAAAAC
VIM-random 87(−)GTTTTTCGCACCNNNCGCTGTATCAATC
RT-VIM-2/fwdTTGTCCGTGATGGTGATGAG
RT-VIM-2/revTGAAAGTGCGTGGAGACTGC
RT-gapA/fwdCGACAAATATGCTGGCCAGG
RT-gapA/revGTAGTAGCGTGAACGGTGGT
  • a The VIM-2-EXP primers were used to clone the blaVIM-2 ORF, which added EcoRI (italicized) and NdeI (underlined) restriction sites at the 5′ end of the gene and XhoI (italicized) and BamHI (underlined) restriction sites after the blaVIM-2 stop codon. VIM-random primers were used to obtain libraries with mutations at selected positions. RT oligonucleotides were used in real-time PCR assays.