TABLE 1.
List of PCR primers used in this studya
| Primer name | Primer sequence (5′ to 3′) | Amplified gene or region |
|---|---|---|
| CR26 (forward) | GGAATTCCATATGGCAGTCGTACCC | def |
| CR27 (reverse) | CCCAAGCTTTTAGTGACCGAACGG | def |
| CR23 (reverse) | TTAAACGCCCCAGCCATG | TD mutant of def |
| CR25 (internal) | ACCGCGCAGGTGTGGTCATCAAT | ID mutant of def |
| CR24 (internal) | ACCACACCTGCGCGGTCCG | ID mutant of def |
a
CR26 and CR27 were used as external primers for the amplification of the def open reading frame along with mutation. GGAATTCCAT in CR26 and CCCAAGCTT in CR27 do not correspond to the genome sequences but have been introduced to incorporate NdeI and HindIII sites at respective 5′ and 3′ ends of the PCR-amplified products. Details of PCR amplification, construction of recombinant plasmid, and generation of PDF mutants were described elsewhere (17).