TABLE 2.
Cleavage efficiency at the p28 site, using substrates of increasing length
| Enzymea | Length (amino acids)b | Mass (kDa)c | Cleavage efficiency (%)d |
|---|---|---|---|
| PstI | 301 | 34.7 | 0 |
| EcoRI | 368 | 40.9 | 5 |
| MscI | 622 | 69.4 | 44 |
| BstBI | 867 | 96.2 | 116 |
| SpeI | 1,160 | 128.0 | 100 |
a
Plasmid pSPNK H1272P (6) was linearized with the enzymes listed; each linearized plasmid was used as template in in vitro-coupled transcription-translation reactions to produce polypeptides of increasing size.
b
Lengths of polypeptides synthesized by using the truncated plasmid templates.
c
Predicted molecular mass of corresponding viral polypeptides.
d
Cleavage efficiency (p28 production) was calculated as described in Materials and Methods; the cleavage efficiency with the longest substrate was arbitrarily set to 100%.