Difference between revisions of "Part:BBa K808030"
(→Usage and Biology) |
(→Usage and Biology) |
||
| Line 8: | Line 8: | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
| + | |||
| + | ====Composition==== | ||
Our fusion protein BBa_K808030 is highly chimeric. It consists of parts from three different bacteria, which are the following (starting from N-terminus to C-terminus): | Our fusion protein BBa_K808030 is highly chimeric. It consists of parts from three different bacteria, which are the following (starting from N-terminus to C-terminus): | ||
| Line 13: | Line 15: | ||
* [https://parts.igem.org/wiki/index.php?title=Part:BBa_K808026 pNB-Esterase 13] (BBa_K808026) is the the catalytical domain of BBa_K808030. It derives from an esterase of ''Bacillus licheniformis''. | * [https://parts.igem.org/wiki/index.php?title=Part:BBa_K808026 pNB-Esterase 13] (BBa_K808026) is the the catalytical domain of BBa_K808030. It derives from an esterase of ''Bacillus licheniformis''. | ||
* [https://parts.igem.org/wiki/index.php?title=Part:BBa_K808027 EstA] (BBa_K808027) serves as a membrane anchor but is an inactive and membrane bound esterase mutant deriving from ''Pseudomonas aeruginosa'' | * [https://parts.igem.org/wiki/index.php?title=Part:BBa_K808027 EstA] (BBa_K808027) serves as a membrane anchor but is an inactive and membrane bound esterase mutant deriving from ''Pseudomonas aeruginosa'' | ||
| + | |||
| + | ====Visualization==== | ||
| + | |||
| + | [[Image:pnb_EstA_ohne_mem.png|500px|left|thumb|The composition and folding of BBa_K808030. Yellow:[https://parts.igem.org/wiki/index.php?title=Part:BBa_K808026 pNB-Esterase 13] (BBa_K808026)after the N-terminal cut-off of PhoA. Gray: inactive catalytical part of [https://parts.igem.org/wiki/index.php?title=Part:BBa_K808027 EstA] (BBa_K808027). Blue: Transmembrane helix of EstA, connecting its inactive catalytical domain and membrane anchor. Red: Anchor domain of [https://parts.igem.org/wiki/index.php?title=Part:BBa_K808027 EstA] (BBa_K808027)]] | ||
| + | |||
Revision as of 20:59, 24 September 2012
RBS-PhoA-His6tag-pNBEst13-Myctag-EstA
RBS-PhoA-His6tag-pNBEst13-Myctag-EstA is a long name for a long protein construct. It is a fusion protein which is secreted into the peri plasma in order to be integrated into the outer membran. When the phoA signal sequence is cut off, the N-terminus is formed by our pNB-Est13. This esterase shows actitivty towards PET and is anchored onto the cell by its inactive EstA membrane anchor.
Annotation: This site is about the construct and composition of our chimeric protein BBa_K808030. For any information about its expression rate or enzyme activity please visit BBa_K808032
Usage and Biology
Composition
Our fusion protein BBa_K808030 is highly chimeric. It consists of parts from three different bacteria, which are the following (starting from N-terminus to C-terminus):
- PhoA (BBa_K808028) is responsible for the peri plasmatic expression of our chimeric protein. This is highly important because once expressed into peri plasma, our construct will self-install into the outer membrane. PhoA is native to E.coli
- pNB-Esterase 13 (BBa_K808026) is the the catalytical domain of BBa_K808030. It derives from an esterase of Bacillus licheniformis.
- EstA (BBa_K808027) serves as a membrane anchor but is an inactive and membrane bound esterase mutant deriving from Pseudomonas aeruginosa
Visualization
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1174
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1700
Illegal NgoMIV site found at 1838
Illegal NgoMIV site found at 2378
Illegal NgoMIV site found at 2747
Illegal AgeI site found at 1245 - 1000COMPATIBLE WITH RFC[1000]