Table 1.
Primers and annealing temperatures used for PCR amplifications
| PCRa | Primer | Primer sequenceb | Size of PCR product (bp) and temp (°C) | Position on plasmid (accession no) |
|---|---|---|---|---|
| silE-silS | sil 1105 | GTT CGT CAT GGT TTC ATG AGCc | 1,915; 58.0 | SATY pMG101, silE 1105–1125 (AF067954) |
| sil 3019 | CTG ACC TTT TTT ATC AGC CTGc | SATY pMG101, silS 3019–2999 (AF067954) | ||
| silS | sil 3019 | CTG ACC TTT TTT ATC AGC CTGc | 741; 58.0 | SATY pMG101, silS 3019–2999 (AF067954) |
| sil 2279 | GCG GGT AAA RAC ATC CTC AATc | SATY pMG101, silS 2279–2299 (AF067954) | ||
| silS-silR | sil 2999 | CAG GCT GAT AAA AAA GGT CAGc | 574; 58.0 | SATY pMG101, silS 2999–3019 (AF067954) |
| sil 3572 | AAC GGC TGG GAT ATC RTS CGc | SATY pMG101, silR 3572–3553 (AF067954) | ||
| silR | sil 3244 | CCC TGA TGG CGA AKG AAG AAc | 329; 58.0 | SATY pMG101, silR 3244–3263 (AF067954) |
| sil 3572 | AAC GGC TGG GAT ATC RTS CGc | SATY pMG101, silR 3572–3553 (AF067954) | ||
| silS-silC | sil 2999 | CAG GCT GAT AAA AAA GGT CAGc | 1,472; 58.0 | SATY pMG101, silS 2999–3019 (AF067954) |
| sil 4470 | CCA GTT GCT GRY TGA AAT AGCc | SATY pMG101, silC 4470–4449 (AF067954) | ||
| silC | sil 4090 | CGG GCT GGC GHA MCT TTT TTGc | 381; 58.0 | SATY pMG101, silC 4090–4111 (AF067954) |
| sil 4470 | CCA GTT GCT GRY TGA AAT AGCc | SATY pMG101, silC 4470–4449 (AF067954) | ||
| silC-silB | sil 4090 | CGG GCT GGC GHA MCT TTT TTGc | 1,942; 58.0 | SATY pMG101, silC 4090–4111 (AF067954) |
| sil 6031 | TCC ATA TCC ATA AAV GGC GATc | SATY pMG101, silB 6031–6011 (AF067954) | ||
| silB-silA | sil 6549 | ATT CAG GGD ATG GAC CCG GTc | 732; 58.0 | SATY pMG101, silB 6549–6568 (AF067954) |
| sil 7280 | CCG GGA TAG CTG GTT TTR ATAc | SATY; pMG101, silA 7280–7260 (AF067954) | ||
| silA-silP | sil 9936 | TGG CGT GGT SAT GCT GAT GTAc | 1,228; 58.0 | SATY pMG101, silA 9936–9956 (AF067954) |
| sil 11163 | TGT TCA CTG GCA TCT TCG GTAc | SATY pMG101, silP 11163–11143 (AF067954) | ||
| silP | sil 11882 | GGC GAT AAG CTC CGC ATC AGAc | 524; 58.0 | SATY pMG101, silP 11882–11902 (AF067954) |
| sil 12405 | TCC ACT TTT TCA AGA CGC TCAc | SATY pMG101, silP 12405–12385 (AF067954) | ||
| RepHIA | RepHIA-F | ACC TTT CTT AAC YTC TTG ATAd | 450; 51.1 | SEMA pR478, 813–793 (BX664015) |
| RepHIA-R | TGT GGT ATC AGT TCW ACA AAC ATd | SEMA pR478, 364–386 (BX664015) | ||
| M13 | M13-F | GTA AAA CGA CGG CCA G | Depending on insert; 50.0 | pCR2.1-TOPO, 391–406 |
| M13-R | CAG GAA ACA GCT ATG AC | pCR2.1-TOPO, 205–221 |
a
The structure of the complete sil operon is depicted in Fig. 2.
b
A = adenine, C = cytosine, G = guanosine, T = thymine, I = inosine; D = G, T or A; H = A, C or T; K = G or T; M = A or C; R = A or G; S = G or C; V = G, C or A; W = A or T; Y = C or T.