TABLE 2.
Restriction patterns of PCR products showing mutations affecting amino acids at various positions compared with blaSHV-1
| Amino acid positiona in blaSHV (codons in blaSHV-1b and mutations in other blaSHV genes) | Endonucleasec | Size(s) (bp) of DNA fragment(s) in blaSHV-1 | Size(s) (bp) of DNA fragment(s) generated by mutation | Genes carrying mutations affecting amino acids at various positions |
|---|---|---|---|---|
| 35 (CTA→CAA) | DdeI | 492, 198, 137 | 629, 198 | blaSHV-2a, blaSHV-11, blaSHV-12, blaSHV-13, blaSHV-15, blaSHV-25 |
| 43 (CGC→AGC) | BglI | 333, 237, 165, 92 | 498, 237, 92 | blaSHV-7, blaSHV-14, blaSHV-18 |
| 54 (GGC→deletion) | EagI, EclXI, Eco521, NotI, XmaIII | 634, 193 | 824d | blaSHV-9, blaSHV-10 |
| 80 (GTG→ATG) | BtsI | 389, 276, 162 | 665, 162 | blaSHV-15 |
| 122 (CTC→TTC) | EcoRI, RsrI | 827 | 433, 394 | blaSHV-21, blaSHV-23 |
| 129 (ATG→GTG) | Hsp92II, NlaIII | 236, 171, 170, 147, 66, 36 | 236, 207, 170, 147, 66 | blaSHV-25 |
| 130 (AGC→GGC) | BstXI, DsaI, EcoT14I, NcoI, StyI | 827 | 414, 410d | blaSHV-10 |
| 140 (GCC→CGG) | AvaII, Eco471, RsrII | 686, 141 | 450, 233, 141d | blaSHV-9, blaSHV-10 |
| BalI, MscI | 375, 264, 188 | 636, 188d | ||
| 140e (GCC→ACN) or 141e (ACC→GCN) | BalI, MscI | 375, 264, 18 | 639, 188 | blaSHV-1, blaSHV-2 |
| 156 (GGC→GAC) | BglI | 333, 237, 165, 92 | 425, 237, 165 | blaSHV-27 |
| TaqI | 827 | 498, 329 | ||
| 158 (AAC→AAG) | MaeII, HpyCH4IV | 506, 321 | 827 | blaSHV-22 |
| BstEII, BstPI, Eco911, Eco0651 | 827 | 506, 321 | ||
| 173 (CTT→TTT) | Bsp143II, BstH2I, HaeII | 434, 192, 116, 85 | 434, 201, 192 | blaSHV-19, blaSHV-20, blaSHV-21 |
| 179 (GAC→AAC) | BstUI | 287, 164, 161, 68, 64, 54, 29 | 316, 164, 161, 68, 64, 54 | blaSHV-8 |
| 179 (GAC→GGC) | Cfr42I, KspI, Sfr303I, SacII, SstII | 827 | 567, 260 | blaSHV-24 |
| 187 (GCC→ACC) | BglI | 333, 237, 165, 92 | 333, 329, 165 | blaSHV-26 |
| 188 (GCG→GGG) | AvaII, Eco47I, SinI | 686, 141 | 596, 141, 90 | blaSHV-23 |
| Eco0109I | 827 | 596, 231 | ||
| 192 (AAG→AAC) | MaeII | 506, 321 | 504, 218, 102d | blaSHV-9, blaSHV-10 |
| 193 (CTG→GTG) | AluI | 306, 153, 151, 91, 84, 42 | 348, 151, 150, 91, 84d | |
| 238 (GGC→AGC) | MaeI, NheI | 971f | 744, 227 | blaSHV-2 to blaSHV-5, blaSHV-2a, blaSHV-7, blaSHV-9, blaSHV-10, blaSHV-12, blaSHV-15, blaSHV-20 to blaSHV-23 |
Amino acid position numbering according to the scheme of Ambler et al. (1).
K. pneumoniae blaSHV-1 gene, strain KPZU-8, GenBank accession number X98100 (14). Mutations are highlighted by boldface type.
Alternative restriction endonucleases are included.
Mutations affecting amino acids at positions 130, 140, 192, and 193 are found in blaSHV-9 (except position 130) and blaSHV-10 genes that also have a deletion of amino acid at position 54; thus, both genes yield an amplimer of 824 bp.
blaSHV-1 and blaSHV-2 genes determined by Barthelemy et al. (cited in reference 6) have threonine (ACN) and alanine (GCN) at amino acid positions 140 and 141, respectively.
Using primer S-8 (this study), as a reverse primer for PCR amplification, instead of that described previously (3).