Difference between revisions of "Part:BBa K2213000"

Revision as of 22:32, 31 October 2017

LacUV5_EutS

The ethanolamine utilisation bacterial microcompartment (BMC) protein, EutS, under control of the LacUV5 inducible promoter. Also contains a bidirectional terminator, RBS and all inducible components of the Lac operon. Thus, this part can be used to synthesize EutS at varying concentrations, depending on the task at hand. Eut S is tagged with His6.

Lac UV5 Promoter

Eut S

Eut S is one of the shell proteins that make up the Ethanolamine utilisation bacterial microcompartment (Eut BMC) in Salmonella spp and other enterobacteria species. It is a hexameric protein, and seem to function as the outer edges of the BMC shell (Held et.al, 2013).

A study conducted by Held et.al (2016) and Choudhary et.al (2012) has shown that Eut S is necessary and sufficient for the successful formation of the Eut BMC. This property was also observed by the CU-Boulder iGEM team in 2016 (http://2016.igem.org/Team:CU-Boulder). While we did not observe the sufficiency of EutS to form microcompartment, our data suggests that EutMN becomes more stable when co-expressed with EutS (see below). This seems to be in line with previous findings on the necessity of EutS for proper BMC formation and further substantiates it.

Usage and Biology

Although it is possible to use this part for EutS expression without further assembly, we do not recommend doing this if the ultimate goal is to produce fully functional Eut BMCs. Despite When forced to produce BMCs, E. coli are placed under a large amount of strain and begin to experience slowed and abnormal growth (see characterisation data below). Therefore, we suggest using a low copy number plasmid eg. pSB4A5 (https://parts.igem.org/Part: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.

Sequence and Features