Difference between revisions of "Part:BBa K2213001"

Revision as of 14:43, 3 September 2017

Tet_EutMN

Ethanolamine Utilisation (Eut) bacterial micro-compartment (BMC) proteins EutM and EutN from E.coli , placed under the inducible Tetracycline promoter. Also contains RBS, terminators and all tetp components – thus this part alone can be used to synthesise EutM and EutN at varying concentrations, relevant to the experimental task.

Usage and Biology

Although it is possible to use this part for EutM and N expression without further assembly, we do not recommend doing this if the ultimate goal is to produce fully functional Eut BMCs. When forced to produce BMCs, E. coli are placed under a large amount of strain and begin to experience slowed and abnormal growth. Therefore, we suggest using a low copy number plasmid (eg. pSB4A5<a href="http://www.humanityhallows.co.uk/igem-manchester-2017-the-path-to-recycling-fertilisers/" target="_blank">http://www.humanityhallows.co.uk/igem-manchester-2017-the-path-to-recycling-fertilisers/</a>, as we have used in our project). By using a low copy number plasmid, cellular stress is minimised, but the experimenter still has the ability to induce BMC formation.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 2190
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 1516

@@ Line 8: / Line 8: @@
 ===Usage and Biology===
-Although it is possible to use this part for EutM and N expression without further assembly, we do not recommend doing this if the ultimate goal is to produce fully functional Eut BMCs. When forced to produce BMCs, <i> E. coli</i>  are placed under a large amount of strain and begin to experience slowed and abnormal growth. Therefore, we suggest using a low copy number plasmid (eg. pSB4A5, as we have used in our project). By using a low copy number plasmid, cellular stress is minimised, but the experimenter still has the ability to induce BMC formation.
+Although it is possible to use this part for EutM and N expression without further assembly, we do not recommend doing this if the ultimate goal is to produce fully functional Eut BMCs. When forced to produce BMCs, <i> E. coli</i>  are placed under a large amount of strain and begin to experience slowed and abnormal growth. Therefore, we suggest using a low copy number plasmid (eg. pSB4A5<a href="http://www.humanityhallows.co.uk/igem-manchester-2017-the-path-to-recycling-fertilisers/" target="_blank">http://www.humanityhallows.co.uk/igem-manchester-2017-the-path-to-recycling-fertilisers/</a>, as we have used in our project). By using a low copy number plasmid, cellular stress is minimised, but the experimenter still has the ability to induce BMC formation.
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