Colicins

Yara Reis Erik Sommer Pascal Krämer Andreas Kühne Kathrin Nußbaum Marika Ziesack

Prey-Strain

• Double-transformation: – Sender for AI-1 and Sender for AI-2  into DH5-a and Top 10 • Check via GFP-Measurement:

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Cloning Strategy for Killer

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

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- will have a Receiver-Vector pSB1A3

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Cloning Strategy for Killer

2. Cloning of the Colicin-cassette behind the receiver-part into Receiver-Vector pSB1A3  restartet cloning this week, that is why still in progress

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Test of colicins effect

2. Try: • Mixing Top10 cell suspension with colicin containing supernatant - Measurement of cell growth curves

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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

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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

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