Table 2.
Primers used in this study
| Primer use | Primer name (sequencea) |
|---|---|
| In-frame deletion of nla28S | |
| nla28S upstream 700 bp | ZS92F (GGATGTCCGTGGGGAGAAACTCCGCAGTTGG) |
| ZS92R (CGACGAGGATGCGGGCTGAGCGCCGTTCCTCTATCACGGGGCC) | |
| nla28S downstream 700 bp | ZS93F (GGCCCCGTGATAGAGGAACGGCGCTCAGCCCGCATCCTCGTCGTG) |
| ZS93R (CCGCTGGCCTCCTCGAAGACGCCTCGCCGC) | |
| Cloning of nla28S, nla28, envZ, and ompR | |
| nla28S-cyt | ZS108F (GCGCCGGGATCCCGCGTCACCTCGCTGCTCAAG) |
| ZS108R (GGGCGGGAATTCTCATGTTCCGACCTTGCGCATTTC) | |
| nla28 | ZS109F (GCGCCGGGATCCAGCTCAGCCCGCATCCTC) |
| ZS109R (GGGCGGGAATTCTCACGACTCGGCCTCCGGGGCCTC) | |
| envZ-cyt | ZS134F (GGCCGCCATATGGCGGATGACCGCACGCTGCTG) |
| ZS134R (GCGCCGAAGCTTCCCTTCTTTTGTCGTGCCCTGC) | |
| ompR | ZS135F (GGCCGCCATATGCAAGAGAACTACAAGATTCTGGTG) |
| ZS135R (GCGCCGAAGCTTTGCTTTAGAGCCGTCCGGTAC) | |
| Site-directed mutagenesis | |
| nla28S-cyt H242A | ZS188F (CCTTCACTCGGCGCGTGGCGGCCGACCTCATCTCCCCGCTGG) |
| ZS188R (CCAGCGGGGAGATGAGGTCGGCCGCCACGCGCCGAGTGAAGG) | |
| nla28S-cyt D386A | ZS189F (GGCCGTGCTGGAGGTCGTGGCCAACGGCATCGGCATGGCGC) |
| ZS189R (GCGCCATGCCGATGCCGTTGGCCACGACCTCCAGCACGGCC) | |
| nla28 D53A | ZS192F (CCTTTGACCTGGTCCTCACGGCCATGGCCATGCCCGAGCCGG) |
| ZS192R (CCGGCTCGGGCATGGCCATGGCCGTGAGGACCAGGTCAAAGG) | |
| qPCR | |
| nla28S (1,030 bp–1,209 bp of nla28S) | ZS155F (CAGTTGTTGCAGGTGAGTGC) |
| ZS155R (GAAGGGCTGGAACAGTGAGG) | |
| rpoD (519 bp–687 bp of rpoD) | ZS156F (CGCGGAAGAGAAGGAAGACG) |
| ZS156R (CTTCTCACCATCCTCGATGC) |
a
Primer sequences are presented in the 5′ to 3′ direction. Restriction endonuclease recognition sites are presented in italics. Mutated codons are presented in bold type.