TABLE 1.
Strains, plasmids, and primers used in this study
| Bacterial strain, plasmid, or primer | Description or sequencea | Sourceb |
|---|---|---|
| Strains | ||
| G1222 | S. dysenteriae serotype 7 type strain | IMVS |
| G1112 | E. coli O7 type strain | NICPBP |
| E. coli BL21(DE3) | F−ompT hsdSB(rB− mB−) gal dcm (DE3) | Novagen |
| E. coli DH5α | F− φ80lacZΔM15 endA recA1 hsdR(rK− mK−) supE44 thi-1 gyrA96 relA1 (lacZYA-argF)U169 | TBC |
| Plasmids | ||
| pET28a+ | T7 expression vector, Kanr | Novagen |
| pLW1048 | pET28a+ containing N-terminally six-histidine-tagged rmlAD7 at the EcoRI/XhoI site | This work |
| pLW1051 | pET28a+ containing C-terminally six-histidine-tagged rmlBD7 at the NcoI/XhoI site | This work |
| pLW1049 | pET28a+ containing N-terminally six-histidine-tagged vioAD7 at the EcoRI/XhoI site | This work |
| pLW1195 | pET28a+ containing N-terminally six-histidine-tagged vioAO7 at the BamHI/HindIII site | This work |
| pLW1155 | pET28a+ containing C-terminally six-histidine-tagged vioBO7 at the NcoI/SalI site | This work |
| Primers | AuGCT | |
| wl-4235 (rmlA-D7F) | 5′-TAGAATTCATGGCTTACTCAGCAGTATG-3′ | |
| wl-4236 (rmlA-D7R) | 5′-TTGGCTCTCGAGTTATTTGTCCTTAAC-3′ | |
| wl-4241 (rmlB-D7F) | 5′-GCAGCCATGGAAATCCTTATTACAG-3′ | |
| wl-4242 (rmlB-D7R) | 5′-GTCCTCGAGTTTCATTTCTCCATACTG-3′ | |
| wl-4243 (vioA-D7F) | 5′-GAGAATTCATGGAAAAGCCAATCTTTGTAAC-3′ | |
| wl-4244 (vioA-D7R) | 5′-GCACTTCTCGAGTCATTTAATCTCCCTAATC-3′ | |
| wl-5702 (vioA-O7F) | 5′-GCAGGATCCATGAACGATAAAACTATTCCAGTAAC-3′ | |
| wl-5703 (vioA-O7R) | 5′-GTCGAAGCTTTCACATCTTACCCAATAATAATTTG-3′ | |
| wl-4704 (vioB-O7F) | 5′-GGTCCCATGGCCTATTTAGATGAAATAC-3′ | |
| wl-4705 (vioB-O7R) | 5′-CGGAAGTCGACCAGATTATCTCCAATAG-3′ |
a
The underlined sequences indicate restriction sites.
b
IMVS, the Institute of Medical and Veterinary Science, Adelaide, Australia; NICPBP, the National Institute for the Control of Pharmaceutical and Biological Products, Beijing, China; TBC, Tianjin Biochip Corporation, Tianjin, China; AuGCT, AuGCT Biotechnology Corporation, Beijing, China.