Table 1.
Primer sequences, restriction enzymes and respective sites, and PCR amplification conditions
| Gene | Primera |
Restriction site | |
|---|---|---|---|
| Type | Sequence (5′-3′) | ||
| sip | Sense | CGCGGATCCCAAGAAACAGATACGACG | BamHI |
| Antisense | GTACGCGGCCGCTTATTTGTTAAATGATACG | NotI | |
| sip-clfA | Sense | CCCGGATCCGTTAAACCAACTCAGACGTCAGTCA | BamHI |
| Overlapping primer 1 | TATTTGTTTTATCGATTTGGAGCCGCCGCCGCCGGAGCCGCCGCCGCCAACTTCATTACCAAGTGCT | ||
| Overlapping primer 2 | GAGCACTTGGTAATGAAGTTGGCGGCGGCGGCTCCGGCGGCGGCGGCTCCCAAATCGATAAAACAAAT | ||
| Antisense | CGCAAGCTTCTCTGGAATTGGTTCAATTTCACCA | HindIII | |
a
The restriction sites are underlined. A 30-bp linker sequence of (G4S)2 flexible bridge (shown in boldface) was added to the overlapping primers for sip-clfA. The melting temperature for all primers was 56°C.