TABLE 1.
Bacterial strains, plasmids, and primers used in this study
| Strains/Plasmids/Primers | Description/Sequence | Source |
|---|---|---|
| E. coli strains | ||
| BL21(DE3) | B; F−ompT hsdSB(rB−mB−) gal dcm (DE3) | Novagen |
| XL1-Blue | RecA1 ebdA1 gyrA96 thi-1 hsdR17 supE44 | Stratagene |
| Plasmids | ||
| pET-22b(+) | Novagen | |
| Primer pairs used for cloning | ||
| KS422/KS273 | 5′-GCATCTCATATGGCTGTTATTATATTTGTTAACG-3′ | |
| WT NmW | 5′-CCGCTCGAGTTTTTCTTGGCCAAAAAACTG-3′ | |
| KS422/KS273 | 5′-GCATCTCATATGGCTGTTATTATATTTGTTAACG-3′ | |
| WT NmY | 5′-CCGCTCGAGTTTTTCTTGGCCAAAAAACTG-3′ | |
| Primer pairs used for the introduction of mutation in SiaDW | ||
| KS350/KS351 | 5′-CTGATCATGACATCAGAAAGTGCGGGATTTCCATATATATTTATG-3′ | |
| E307A | 5′-CATAAATATATATGGAAATCCCGCACTTTCTGATGTCATGATCAG-3′ | |
| KS370/KS371 | 5′-ATCTCGCGTTGCTGTAGGTGTTTATGCAACTAGCTTATTTG-3′ | |
| S972A | 5′-CAAATAAGCTAGTTGCATAAACACCTACAGCAACGCGAGAT-3′ | |
| Primer pairs used for the introduction of mutation in SiaDY | ||
| AR11/AR12 | 5′-ATACAGATATCCTAATCATGACATCTCAAAGCGCAGGCTTTGGTTATATAT-3′ | |
| E307A | 5′-atatataaccaaagcctgcgctttgagatgtcatgattaggatatctgtat-3′ | |
| KS370/KS371 | 5′-ATCTCGCGTTGCTGTAGGTGTTTATGCAACTAGCTTATTTG-3′ | |
| S972A | 5′-CAAATAAGCTAGTTGCATAAACACCTACAGCAACGCGAGAT-3′ | |
| Primer pairs used for the generation of SiaDW truncations | ||
| KS422/KS421 | 5′-GCATCTCATATGGCTGTTATTATATTTGTTAACG-3′ | |
| CΔ639 | 5′-CCGCTCGAGGCTGCGCGGAAGAATAGTG-3′ | |
| AR2/KS273 | 5′-GCATCTCATATGTTTAATAACGTATCATTATCGTC-3′ | |
| NΔ398 | 5′-CCGCTCGAGTTTTTCTTGGCCAAAAAACTG-3′ | |
| AR1/KS273 | 5′-GCATCTCATATGACTGATGATAATTTAATACCTAT-3′ | |
| NΔ562 | 5′-CCGCTCGAGTTTTTCTTGGCCAAAAAACTG-3′ | |
| AR9/AR273 | 5′-GCATCTCATATGAAATATTCTTATAAATATATCTA-3′ | |
| NΔ609 | 5′-CCGCTCGAGTTTTTCTTGGCCAAAAAACTG-3′ | |
| AR10/KS273 | 5′-GCATCTCATATGTCTTGGGAACTTATTCGTGCCTC-3′ | |
| NΔ639 | 5′-CCGCTCGAGTTTTTCTTGGCCAAAAAACTG-3′ | |
| KS433/KS273 | 5′-GCATCTCATATGGGTAAGCGTTCGATGGATG-3′ | |
| N-Δ676 | 5′-CCGCTCGAGTTTTTCTTGGCCAAAAAACTG-3′ | |
| KS434/KS273 | 5′-GCATCTCATATGTCACTGAAAAGTAATGTAGTTG-3′ | |
| NΔ729 | 5′-CCGCTCGAGTTTTTCTTGGCCAAAAAACTG-3′ | |
| KS435/KS273 | 5′-GCATCTCATATGAATATCGAAGCATTTCTAAAACC-3′ | |
| NΔ777 | 5′-CCGCTCGAGTTTTTCTTGGCCAAAAAACTG-3′ | |
| Primer pairs used for the generation of SiaDY truncations | ||
| KS422/KS421 | 5′-GCATCTCATATGGCTGTTATTATATTTGTTAACG-3′ | |
| CΔ639 | 5′-CCGCTCGAGGCTGCGCGGAAGAATAGTG-3′ | |
| AR2/KS273 | 5′-GCATCTCATATGTTTAATAACGTATCATTATCGTC-3′ | |
| NΔ398 | 5-´CCGCTCGAGTTTTTCTTGGCCAAAAAACTG-3′ | |
| AR1/KS273 | 5′-GCATCTCATATGACTGATGATAATTTAATACCTAT-3′ | |
| NΔ562 | 5′-CCGCTCGAGTTTTTCTTGGCCAAAAAACTG-3′ | |
| KS433/KS273 | 5′- GCATCTCATATGGGTAAGCGTTCGATGGATG-3′ | |
| NΔ676 | 5-´CCGCTCGAGTTTTTCTTGGCCAAAAAACTG-3′ | |
| KS434/KS273 | 5′-GCATCTCATATGTCACTGAAAAGTAATGTAGTTG-3′ | |
| NΔ729 | 5-´CCGCTCGAGTTTTTCTTGGCCAAAAAACTG-3′ | |