TABLE 3.
Production of mutant constructs: internal mutations
| Constructsa | PCRb
|
||
|---|---|---|---|
| 1st amplification
|
2nd amplification
|
||
| Template | Primers | Primers | |
| pvTRΔAP1 | pvTR | 802/712 | 802/729 |
| pvTR | 713/729 | ||
| pvTRΔE1 | pvTR | 802/714 | 802/729 |
| pvTR | 715/729 | ||
| pvTRΔE2 | pvTR | 802/716 | 802/729 |
| pvTR | 717/729 | ||
| pvTRΔCAATx | pvTR | 802/702 | 802/729 |
| pvTR | 703/729 | ||
| pvTRΔCAATy | pvTR | 802/704 | 802/729 |
| pvTR | 705/729 | ||
| pchTRΔCAATx | pchTR | M165/M167 | M165/M166 |
| pchTR | M168/M166 | ||
| pchTRΔCAATy | pchTR | M165/M169 | M165/M166 |
| pchTR | M170/M166 | ||
a
The Δ symbol means that the following site specified is mutated. E1 and E2 correspond to E-box 1 and E-box 2, respectively.
b
Primer sequences are described in Table 2. Primers which comprise desired mutations are in bold.