Table 2.
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) |
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.