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Killing by Colicin 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
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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