Notebook : August 26

summary

• started the Restriction digestion and ligation to get the BioBrick fnr into plasmid PSB1C3 by using restrictin ensyme EcoRI and PstI.

• Sent an E-mail to M. Nesbeth from UCL for requesting the Biobrick BBa_K239005 which was created by iGEM UCL team in 2009

• Designed oligonucleotides for the gene coding for BphR2 (regulator, sensitive for PCBs), Transcriptional regulator of Pseudomonas oleovorans /pseudoalcaligenes group

lab work

• A.aero/anaerobic regulation system
  
  • 1.BioBrick promotor fnr(repressor and activator) in plasmid PSB1C3

Digestion for promoter fnr(repressor and activator) in PSB1C3

2 enzymes SPE I and PST I

For promoter fnr(repressor) :

For promoter fnr(activator)nirB clone 6:

For promoter fnr(activator)nark clone 6 :

For promoter fnr(activator)nirB clone 7 :

For promoter fnr(activator)narG clone 6 :

For promoter fnr(activator)nark clone 6 :

For promoter fnr(activator)nirB clone 6 :

For promoter fnr(activator)nark Digested SPE I :

Ligation

After 3h of digestion, we mixed the digestion products:

Then we performed a precipitation by ethanol in order to inactivate the enzymes. And suspended the deposit by:

The incubation was during 1H30.

• B.PCBs sensor system
  
  • 1.BioBrick BphR2(regulator) in plasmid PSB1C3
    

Using software gene manager to find the oligopeptide for amplification of BphR2.