Difference between revisions of "Part:BBa K3482004"

Revision as of 16:50, 26 October 2020

IPTG-inducible miniColicin toxin

This part allows for IPTG inducible expression of the DNAase miniColicin E2 in E. coli. The degradation of DNA , which leads to cell death, can therefore be induced by IPTG.

Biology

Colicin E2 is an endonuclease that cuts on both single and double-stranded DNA. It has no specific cutting site. Colicins are toxins that can be produced by E.coli and closely related bacteria.

Usage

We used that part to test if the expression of the toxin work in our E.coli nissle 1917 strain and if it was sufficient do provoc cell death

Characterization

Functional killswitch assay with IPTG and aTc gradients on agar plate

E.coli Nissle 1917 were transformed with a plasmid containing the part BBa K3482004 and the part BBa_K3482014 (IM2 antitoxin part) and plated with a gradient of aTc and IPTG on agar plate. The plate shows strong activity of the IM2 antitoxin with aTc induction, whereas IPTG induction promotes production of the MiniColicin E2 toxin, resulting in a number of surviving cells (probable mutants) proportional to the dilution of the plated culture.

Sequence and Features