Colicins

Yara ReisErik Sommer

Pascal KrämerAndreas Kühne

Kathrin Nußbaum Marika Ziesack

Killing by Colicin 208.05.23

• Double-transformation: – Sender for AI-1 and Sender for AI-2

into DH5-a and Top 10

• Check via GFP-Measurement:

Prey-Strain

Killing by Colicin 308.05.23

1. Cloning of the receiver-part into the pSB1A3-Vector- receiver-part via PCR from T9002:

- will have a Receiver-Vector

Cloning Strategy for Killer

pSB1A3

Rec

Killing by Colicin 408.05.23

2. Cloning of the Colicin-cassette behind the receiver-part into Receiver-Vector

restartet cloning this week, that is why still in

progress

Cloning Strategy for Killer

pSB1A3

Rec

Co

l

Killing by Colicin 508.05.23

Test of colicins effect

2. Try:• Mixing Top10 cell suspension with colicin containing

supernatant- Measurement of cell growth curves

Killing by Colicin 608.05.23

Test of colicins effect

2. Try:• Results:

- no significant difference between normal growthcurve and that with colicin supernatant

- perhaps the cells died because of mytomycin instead of colicin

• Advances:- try other methods to stress colicin producing

cells- measure decreasing GFP-expression of cells

that shall be killed by colicin instead of OD

Killing by Colicin 708.05.23

Future

• Cloning of colicin behind receiver verify and visualize

• Test of colicin effect on e.coli strains and aditionally on different eukaryotic cell lines

• Purify colicin-proteins via His-Tag for modeling parameters