Difference between revisions of "Part:BBa K3697002"
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This part is the expression cassette for expressing manP in B. subtilis. It includes the natural RBS, Promoter, and coding sequence for the manP gene found in Bacillus Subtilis 168. Because of this, this cassette can be used in B. subtilis to produce this transporter protein at normal levels. | This part is the expression cassette for expressing manP in B. subtilis. It includes the natural RBS, Promoter, and coding sequence for the manP gene found in Bacillus Subtilis 168. Because of this, this cassette can be used in B. subtilis to produce this transporter protein at normal levels. | ||
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| + | [1] Wenzel, M. and Altenbuchner, J. (2015) Development of a markerless gene deletion system for Bacillus subtilis based on the mannose phosphoenolpyruvate‐dependent phosphotransferase system. Microbiology (United Kingdom), 161(10), 1942–1949. | ||
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| + | <!-- Add more about the biology of this part here> | ||
| + | ===Usage and Biology=== | ||
The manP gene in B. Subtilis is a part of the mannose phosphotransferase system (PTS). The mannose PTS is primarily encoded by two genes manA and manP. ManP encodes for the specific transporter used by B. subtilis to uptake mannose into the cell and phosphorylate it, whereas manA encodes for mannose-6-isomerase which converts mannose into fructose [1]. These two genes work in tandem to take mannose up into the cell and convert it into a form that is usable in the cell. | The manP gene in B. Subtilis is a part of the mannose phosphotransferase system (PTS). The mannose PTS is primarily encoded by two genes manA and manP. ManP encodes for the specific transporter used by B. subtilis to uptake mannose into the cell and phosphorylate it, whereas manA encodes for mannose-6-isomerase which converts mannose into fructose [1]. These two genes work in tandem to take mannose up into the cell and convert it into a form that is usable in the cell. | ||
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Unfortunately, due to limitations in lab space due to the pandemic the team was not able to independently verify manP as a negative selection marker, but its use as a negative selection marker is well documented in the 2015 paper by Marian Wenzel and Josef Altenbuchner which can be found at this link: https://www-microbiologyresearch-org.stanford.idm.oclc.org/content/journal/micro/10.1099/mic.0.000150a) | Unfortunately, due to limitations in lab space due to the pandemic the team was not able to independently verify manP as a negative selection marker, but its use as a negative selection marker is well documented in the 2015 paper by Marian Wenzel and Josef Altenbuchner which can be found at this link: https://www-microbiologyresearch-org.stanford.idm.oclc.org/content/journal/micro/10.1099/mic.0.000150a) | ||
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Revision as of 00:33, 24 October 2020
manP expression cassette
This part is the expression cassette for expressing manP in B. subtilis. It includes the natural RBS, Promoter, and coding sequence for the manP gene found in Bacillus Subtilis 168. Because of this, this cassette can be used in B. subtilis to produce this transporter protein at normal levels.
[1] Wenzel, M. and Altenbuchner, J. (2015) Development of a markerless gene deletion system for Bacillus subtilis based on the mannose phosphoenolpyruvate‐dependent phosphotransferase system. Microbiology (United Kingdom), 161(10), 1942–1949.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 129
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 722
Illegal AgeI site found at 372
Illegal AgeI site found at 466
Illegal AgeI site found at 2132 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 272
Illegal SapI site found at 1294