TABLE 1.
PCR primers used for construction of mutant PhaJAc molecules
| Primer direction | Primer | Amino acid substitution | Sequencea |
|---|---|---|---|
| Forward | fS62A | Ser-62 to Ala | 5′-CTCTTCGCCGGGCTGCTGGGCCAGCAGTTG-3′ |
| fL65I | Leu-65 to Ile | 5′-CTCTTCTCCGGGCTGATCGGCCAGCAGTTG-3′ | |
| fL65V | Leu-65 to Val | 5′-CTCTTCTCCGGGCTGGTGGGCCAGCAGTTG-3′ | |
| fL65A | Leu-65 to Ala | 5′-CTCTTCTCCGGGCTGGCGGGCCAGCAGTTG-3′ | |
| fL65G | Leu-65 to Gly | 5′-CTCTTCTCCGGGCTGGGGGGCCAGCAGTTG-3′ | |
| fV130A | Val-130 to Ala | 5′-GGGGAAGCCGTGGCCAAGCTGCCTTAA-3′ | |
| fV130G | Val-130 to Gly | 5′-GGGGAAGCCGTGGGCAAGCTGCCTTAA-3′ | |
| Reverse | rS62/L65 | Ser-62 to Ala, Leu-65 to Ile, Val, Ala, or Gly | 5′-GCTGGCGAGCAGCATGCCGTGGACTATGGG-3′ |
| rV130 | Val-130 to Ala or Gly | 5′-CGTCACGGCGAGGGCGCCGCCTTGGGT-3′ |
a
The underlined sequences are the mutation sites.