TABLE 2.
Primers used in this work
| Purpose | Name | Sequence,a 5′ → 3′ | Description |
|---|---|---|---|
| pilAY32F construction | rLC43F | AGAGCAGGTGAAGGAAGGGAGTTT | Amplifies the last 500 bp of GSU1495 (pilR) with the kanamycin cassette |
| rLC46R | tgacgattttcgtcactggctcctctTGGATCCCCCGGGCTGCAGGAATTCG | ||
| rLC47F | cgaattcctgcagcccgggggatccaAGAGGAGCCAGTGACGAAAATCGTCA | Amplifies pilA | |
| rLC51R | CTATTGCGCACAATGGCTATTCCCTGCATTGCGA | ||
| Plasmid pLC9 construction | Y32F_Fwd | CAGTTCTCGGCGTATCGTGTCAAGGCATTCAACAGCGCGGCGTCAAGCG | Introduces the Tyr-32-Phe mutation |
| Y32F_Rev | CGCTTGACGCCGCGCTGTTGAATGCCTTGACACGATACGCCGAGAACTG |
a
Annealing nucleotides are in lowercase, BsmI restriction sites are italicized and underlined, and base pairs replaced by site-directed mutagenesis are in boldface font.