TABLE 2.
Oligonucleotides used in these studies
| Primer | Sequence (5′→3′)a | Description |
|---|---|---|
| jf031912.1 | TCCGAGCTCGAGATCCTTGTCACTTGCGTTATTAATGAATAAG (XhoI site underlined) | yghJ forward cloning primer |
| jf031912.2 | CCCAAGCTTCGAATTCTCGGCAGACATCTTATGCTC (HindIII site underlined) | yghJ reverse cloning primer |
| jf082312.1 | GCTGATCTGGCATGATGTCGGTCATAACGCC (GAA→GAT mutation underlined) | Site-directed mutagenesis (E1309D) forward |
| jf082312.2 | GGCGTTATGACCGACATCATGCCAGATCAGC | Site-directed mutagenesis (E1309D) reverse |
| jf082312.3 | GGCTGATCTGGCATGCAGTCGGTCATAACGC (GAA→GCA mutation underlined) | Site-directed mutagenesis (E1309A) forward |
| jf082312.4 | GCGTTATGACCGACTGCATGCCAGATCAGCC | Site-directed mutagenesis (E1309A) reverse |
| jf112612.1 | ACGACTGGCTGATCTGGGCTGAAGTCGGTCATAACG (CAT→GCT mutation underlined) | Site-directed mutagenesis (H1308A) forward |
| jf112612.2 | CGTTATGACCGACTTCAGCCCAGATCAGCCAGTCGT | Site-directed mutagenesis (H1308A) reverse |
| jf112612.3 | CTGGCATGAAGTCGGTGCTAACGCCGCAGAAACG (CAT→GCT mutation underlined) | Site-directed mutagenesis (H1312A) forward |
| jf112612.4 | CGTTTCTGCGGCGTTAGCACCGACTTCATGCCAG | Site-directed mutagenesis (H1312A) reverse |
| jf101712.1 | GACTGGCTGATCTGG(−66)AACGTGCTGGCGCTG | Mutagenesis primer to construct 22-amino-acid internal deletion of YghJ residues H1308 to N1329 |
| jf101712.2 | CAGCGCCAGCACGTT(−66)CCAGATCAGCCAGTC | Reverse complement of jf101712.1 |
| jf101812.3 | CCGAAGAAGAACCTGAATGC | Forward sequencing primer yghJ from nt 3616 to 3635 |
| jf101812.4 | TTACCGTTGGATTCAGCACA | Reverse sequencing primer yghJ from nt 4319 to 4300 |
a
(−66) indicates the position of the 66-nucleotide in-frame deletion in yghJ extending from nucleotide (nt) 3904 to 3969.