EPF-Lausanne/9 September 2009

9 September 2009

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Wet Lab

Characterization

	With Atc	Without Atc
With Trp	1/2 Trp + 1/2 Atc 1 Trp + 1/2 Atc 1/2 Trp + 1 Atc 1 Trp + 1 Atc	1/2 Trp 1 Trp 3/2 Trp
Without Trp	1/2 Atc 1 Atc 3/2 Atc	Without Atc nor Trp

Results of the characterization

For the RBS :

RBS was our negative control. We can see that there is no RFP fluorescence (because of course RBS has no RFP gene).

For LacI-RFP 1 + IPTG :

LacI-RFP 2 + IPTG show the same tendancy.

For Read Out 1

RO1#1 + 0.5 TRP :

RO1#2 + 0.5 TRP shows the same tendancy.

RO1#1 + 1 TRP :

Again, RO1#2 clone has the same behaviour.

RO1#1 without TRP:

Clone #2 has the same behaviour.

RO1#3 clones doesn't follow these tendancy. It actually seems that this clone isn't working at all. It is possible a spontaneous mutation occured in some place... On all different conditions, RO1#3 clones show a flat curve, like RBS. It doesn't express RFP which means either our construct wasn't inserted, either there has been a mutation in it. For example + 1 TRP :

For Read Out 2

RO2#1 without TRP :

RO2#3 has exactly the same curve shape :

RO2#1 + 0.5 TRP :

RO2#3 has a more "normal" shape :

RO2#1 + 1 TRP :

RO2#3 is the same :

RO2#1 + 1.5 TRP :

RO2#1 + 0.5 TRP + 0.5 ATC :

RO2#1 + 0.5 TRP + 1 ATC :

RO2#1 + 1 TRP + 0.5 ATC :

RO2#1 + 1 TRP + 1 ATC :

RO2#1 + 0.5 ATC :

RO2#1 + 1 ATC :

RO2#1 + 1.5 ATC :

Clone RO2#2 is not working, it has always a flat curve like this (for ex +0.5 ATC):

People in the lab

Mélanie, Caroline, Basile, Nicolas