Table 2. Incorporation of [U-14C]Gal mediated by WciS–His6 into the GelK product.
The 12K supernatants were prepared from E. coli BL21 (DE3)/ΔgalU cells expressing WciS–His6 (pET-wciS) or not (control vector, pET23a+). Activity corresponding to 50 μl of GT preparation (approx. 300 μg of total proteins) was expressed as pmol of [U-14C]Glc incorporated per h from UDP-[U-14C]Glc on to the glycolipid fraction prepared from a E. coli BL21 (DE3) strain expressing GelK (pET-gelK) or harbouring the corresponding control vector (pET29b, see the Materials and methods section for further details).
| Plasmid expressed | pET29b glycolipid | pET-gelK glycoloipid | UDP-[U-14C]Gal | [14C]Gal incorporated into the acceptor (pmol/h) |
|---|---|---|---|---|
| pET23a+ | − | − | + | <3 |
| pET-wciS | − | − | + | <3 |
| pET23a+ | + | − | + | <3 |
| pET-wciS | + | − | + | 154±31 |
| pET23a+ | − | + | + | <3 |
| pET-wciS | − | + | + | 301±54 |