Table 2. Restriction enzyme digestion patterns predicted for reverse transcription–polymerase chain reaction (RT–PCR) products for each guinea-pig CD1 (gpCD1) isoform.
| Guinea-pig CD1 isoforms | |||||||
|---|---|---|---|---|---|---|---|
| Restriction endonuclease | b1 | b2 | b3 | b4 | c1 | c2 | c3 |
| None | 477 | 477 | 477 | 477 | 477 | 477 | 477 |
| SalI | – | – | – | – | 430/47 | 430/47 | 430/47 |
| StyI | 247/230 | – | 391/86 | – | – | 292/185 | – |
| StuI | – | 394/83 | – | – | – | 259/218 | – |
| XbaI | – | – | – | 246/231 | – | – | – |
| XhoI | – | – | – | – | 420/57 | 420/57 | – |
| DraI | – | – | – | – | – | – | 295/182 |
| KpnI | – | 243/234 | – | – | – | – | – |
| EcoRV | – | 334/143 | 288/189 | – | – | – | – |
| HindIII | – | 313/164 | 313/164 | – | – | 313/164 | – |
Transcripts corresponding to all isoforms of gpCD1 genes could be amplified by PCR with equal efficiency using the universal gpCD1 primers to yield PCR products of 477 bp. From the nucleotide sequences of each isoform, digestion patterns for PCR products corresponding to each isoform of gpCD1were predicted as shown (numbers indicate fragment length in base pairs). For example, DraI digests only the PCR product from gpCD1c3 (295 and 182 bp fragments), and will not digest the PCR product corresponding to other known gpCD1 isoforms. These predicted restriction enzyme digestion patterns were each confirmed by actual digestion of PCR products generated by using plasmids containing cDNA inserts for each isoform of all seven known group 1 gpCD1 genes as templates.