Difference between revisions of "Part:BBa K2333435"

Revision as of 23:45, 1 November 2017

pBad mf-Lon

This is an arabinose-inducible E. coli optimized mf-Lon construct. Arabinose inducibility is provided by BBa_I0500, William and Mary 2017 iGEM found early on that this part had leaky expression, and thus did not perform any high-quality characterization of this part.

Usage and Biology

This part contains mf-Lon under the control of the inducible pBad promoter. The mf-Lon protease specifically targets protein degradation tags (pdts) with various corresponding to varying degradation rates. This arabinose-inducible mf-Lon construct was used in tandem with aTc-inducible pdt reporter constructs to obtain gene expression speed measurements.

Characterization

A concentration of .1mM arabinose was sufficient to create degradation of mScarlet-I and sfGFP [http://2017.igem.org/Team:William_and_Mary/Parts parts], 1mM arabinose increased degradation but lead to high amounts of cell death. .01mM Arabinose generally increased expression. Sequence and Features

References

[1] Cameron DE, Collins JJ. Tunable protein degradation in bacteria. Nature Biotechnology. 2014;32(12):1276–1281.

[2] Torella JP, Boehm CR, Lienert F, Chen J-H, Way JC, Silver PA. Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly. Nucleic Acids Research. 2013;42(1):681–689.