Death gene

The confirmatory experiment of Death Gene

The confirmation experiment of Death Gene

The confirmatory experiment of Death gene.pdf
This part we use R0010 promoter ,it is to say in the absence of LacI protein and CAP protein, it promotes transcription,in the presence of LacI protein and CAP protein, it inhibits transcription. When induced by IPTG, the protein expression begin. Experimental procedure: Step 1: Recovery J04500+K117000+B0015,shaking it in 37℃ for 12 to 16h at 150rpm. Step 2: Measure the OD600 of bacterial liquid. Step 3: Use 50ul bacterial liquid which has been diluted times to coat plates.Set IPTG(none or 1mM) control. Step 4: Add 5ul bacterial liquid which has been diluted times to 5ml liquid Luria-Bertani medium. Set IPTG(none or 1mM) control. After cultivate in 37℃ shaking(150rmp) for 12 to 16h,measure the OD600 of bacterial liquid . Step 5: Use 50ul bacterial liquid(from step 4) which has been diluted times to coat plates. Set IPTG(none or 1mM) control. Step 6: Compare bacterial liquid concentration and petri dishes on the growth of the colony to judge lethal gene’s effects .Compare the bacterial concentration and Petri dish colony growth to judge the effect of lethal gene. Judging from the OD600 of bacterial liquid and the growth state of bacteria in Petri dish to see whether the death gene work or not. Fig 1. The experimental procedure of Death gene’s confirmatory experiment The data of Sweet gene’s confirmatory experiment version 1.0 Fig 2. The growth state of bacteria in Petri dish (Notes: d/e/f means the bacterial liquid were diluted 104/105/106times,A/B means the concentration of IPTG none/1mM ) Fig 3. .The comparation of colony growth (Notes: The left has IPTG of 1mM and the right has none)