Part:BBa_K2235007
OmpR promoter regulating the expression of sialidase enzyme
OmpR is an osmotic pressure promoter. It can sense changes in for example sucrose concentration. A higher concentration of sucrose will lead to a higher expression of the gene after the OmpR promoter. In this biobrick the gene after OmpR is an enzyme called sialidase. This enzyme cuts sialic acid (that's for example found in mucus). The sialidase produced by this biobrick has a HIS-tag on it to make it easy to purify.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 189
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 636
Illegal NgoMIV site found at 711
Illegal NgoMIV site found at 801 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1181
Usage and biology
OmpR is an already well characterized biobrick (BBa_R0082). It is a promoter that takes advantage of the OmpR/EnvZ two-component system, which is utilized by many bacteria, to sense the change in osmotic pressure. We connected this promoter to the coding sequence for the enzyme sialidase. This enzyme hydrolyzes glycosidic linkages of terminal sialic acid residues in glycoproteins. This composite biobrick can be used to produce sialidase when the osmotic pressure in the environment around the bacteria increases.
Important parameters
| Experiment | Characteristic | Result |
|---|---|---|
| Expression | ||
| Compatibility | E. coli TOP10
E. coli ΔEnvZ | |
| Promoter | OmpR | |
| Optimal temperature | 37 °C | |
| Number of amino acids | XXX | |
| Molecular weight | XXXnbsp;kDa | |
| Purification | ||
| Tag | C-terminal 6xHis | |
| Sialidase expression | ||
| SDS-PAGE | Sialidase expression seen in some tests | |
| Osmotic pressure detection | ||
| Sucrose | Works | |
| NaCl | Non-conclusive data |
| None |