TABLE 2.
Primers used for generation of deletion plasmids
| Inactivated gene | Primer (sequence) used for amplification of the gene + flanking sequences | Primer (sequence) used for aac(3)IV amplificationa |
|---|---|---|
| hutF | oTN203 (CAAATTAAGCTTTTCGCCTTCCCCGGTTTCGT; HindIII restriction site is underlined) | oTN207 (TGTGGCCGCACTCGCGGAGGTACTGGAGGGCCTGGCGTGATTCCGGGGATCCGTCGACC) |
| oTN204 (TATTATTCTAGACCACCG TCAGCGAGTTGGCC; XbaI restriction site is underlined) | oTN208 (TGCTCTCCTTCGTCGGCCGTGCCGGTAGCGGCCGGGTCATGTAGGCTGGAGCTGCTTC) | |
| rapK | oTN247 (GACTGTAAGCTTCACCTCACGAACGTCTTC; HindIII restriction site is underlined) | oTN253 (TTCTTCCTGGTACGCGCCAGGCAACGGCGGGGAAGGGTGATTCCGGGGATCCGTCGACC) |
| oTN252 (GAGTCCTCTAGAGATAGGTTTCCATCGGCAG; XbaI restriction site is underlined) | oTN254 (GCTCATCGGTGGTCCTTCCGGGTCAACTCGGCGGGGTCATGTAGGCTGGAGCTGCTTC) |
a
The priming sites of the aac(3)IV disruption cassette are in boldface.