TABLE 4.
ECACYC synthesis in E. coli K-12 strains containing null mutations in the wzyE, wzzE, and wzxE genes
| Enzyme | Strain | Relevant genotype | Synthesis of ECAPGa | Concn of ECACYCb (nmol/mg protein) |
|---|---|---|---|---|
| Polymerase (WzyE) | W3110 | Wild type | + | 3.69 |
| PR4237 | Like W3110, but wecG::Tn10 | − | ND | |
| PR4214 | wecG::Tn10/pRL163 (wild-type wecG under the control of the pBAD promoter) | + | 5.14 | |
| PR4216 | wecG::Tn10 wzyE::gm/pRL163 (wild-type wecG under the control of the pBAD promoter) | − | ND | |
| PR4217 | wecG::Tn10 wzyE::gm/pRL164 (wild-type wzyE and wecG under the control of the same pBAD promoter) | + | 19.7 | |
| PR4224 | wecG::Tn10/pBAD102/D/lacZ | − | ND | |
| Chain length regulator | PR4220 | Like W3110, but wzzE::km | + | ND |
| (WzzE) | PR4219 | wecG::Tn10 wzzE::km/pRL163 (wild-type wecG under the control of the pBAD promoter) | + | ND |
| PR4221 | wzzE::km/pCA53 (wild-type wzzE) | + | 2.97 | |
| PR4226 | wzzE::km/pBR322 | + | ND | |
| Translocase (WzxE) | Sθ874 | Δ(wzxO16wzxC) | + | 2.92 |
| PR4210 | Like Sθ874, but wecA::km wzxE::cm/pRL147 (wild-type wecA under the control of the pBAD promoter), pRL165 (wild-type wzxE under the control of the lac promoter) | + | 2.84 | |
| PR4236 | Like Sθ874, but wecA::km wzxE::cm/pRL147 (wild-type wecA under the control of the pBAD promoter), pRL166 (wild-type wzxC under the control of the lac promoter) | + | 1.51 | |
| PR4233 | Like Sθ874, but wecA::km wzxE::cm/pRL147 (wild-type wecA under the control of the pBAD promoter), pRL169 (wild-type wzxO16 under the control of the lac promoter) | + | 1.92 | |
| PR4225 | Like Sθ874, but wecA::km wzxE::cm/pACYC184 | − | ND | |
| PR4223 | Like W3110, but wecG::Tn10 wzxE::cm/pRL163 (wild-type wecG under the control of the pBAD promoter) | + | 3.9 |
a
Determined by passive hemagglutination assay as described in Materials and Methods.
b
Amount in cell extracts. The preparation of cell extracts and the quantification of ECACYC are described in detail in Materials and Methods. ND, not detected.