Difference between revisions of "Part:BBa K5136059"
| Line 7: | Line 7: | ||
The rhamnosyltransferase (RhlB) comes from <i>Pseudomonas aeruginosa</i>, is capable of catalyzing the reaction between HAA and dTDP-l-rhamnose to form mono-rhamnolipid. (1) The hydropathy plot of the RhlB suggested that the protein is membrane-anchored via its N-terminal, crossing the membrane and exposing its domains on both sides of the membrane.<br> | The rhamnosyltransferase (RhlB) comes from <i>Pseudomonas aeruginosa</i>, is capable of catalyzing the reaction between HAA and dTDP-l-rhamnose to form mono-rhamnolipid. (1) The hydropathy plot of the RhlB suggested that the protein is membrane-anchored via its N-terminal, crossing the membrane and exposing its domains on both sides of the membrane.<br> | ||
| − | ===Usage | + | ===Usage=== |
We used <partinfo>BBa_K081005</partinfo> to construct the expression system and obtained the composite <partinfo>BBa_K5136232</partinfo>, which is assembled on the expression vector pSB1C3 by standard assembly. The constructed plasmid was transformed into <i>E. coli</i> DH5α, then the positive transformants were selected by chloramphenicol and confirmed by colony PCR and sequencing.<br> | We used <partinfo>BBa_K081005</partinfo> to construct the expression system and obtained the composite <partinfo>BBa_K5136232</partinfo>, which is assembled on the expression vector pSB1C3 by standard assembly. The constructed plasmid was transformed into <i>E. coli</i> DH5α, then the positive transformants were selected by chloramphenicol and confirmed by colony PCR and sequencing.<br> | ||
Revision as of 06:16, 29 September 2024
rhlB
Transferase activity, transferring hexosyl groups.
Biology
The rhamnosyltransferase (RhlB) comes from Pseudomonas aeruginosa, is capable of catalyzing the reaction between HAA and dTDP-l-rhamnose to form mono-rhamnolipid. (1) The hydropathy plot of the RhlB suggested that the protein is membrane-anchored via its N-terminal, crossing the membrane and exposing its domains on both sides of the membrane.
Usage
We used BBa_K081005 to construct the expression system and obtained the composite BBa_K5136232, which is assembled on the expression vector pSB1C3 by standard assembly. The constructed plasmid was transformed into E. coli DH5α, then the positive transformants were selected by chloramphenicol and confirmed by colony PCR and sequencing.
Reference
1. Chong, H.; Li, Q., Microbial production of rhamnolipids: opportunities, challenges and strategies. Microb Cell Fact 2017, 16 (1), 137.
2. Ochsner, U. A.; Fiechter, A.; Reiser, J., Isolation, characterization, and expression in Escherichia coli of the Pseudomonas aeruginosa rhlAB genes encoding a rhamnosyltransferase involved in rhamnolipid biosurfactant synthesis. J Biol Chem 1994, 269 (31), 19787-19795.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 1138
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 31
Illegal NgoMIV site found at 865 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 381