Difference between revisions of "Team:ShanghaiTechChina B/Description"
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Revision as of 01:14, 20 October 2016
Description
This year, we mainly tested two different kinds of killer genes, and turned to characterize them under different expression circuits.
During our experiment, we compared the property of killer red with gene E, both of them were linked to the same T7 promoter and RBS. At the same time, we tested the efficiency of gene E in different plasmid: pet28a and pBV220.
Induced by IPTG, E. coli transformed with killer red and gene E showed different behaviors. Under green laser, red fluorescence could be observed in induced E. coli which contained killer red. We got perfect image of red fluorescent by using confocal microscopy. By experiment, killer red could not be activated by light of shaker, but easily activated by green laser of fluorescence microscope. Using spread-plant method we tested the efficiency of killer red and got positive results. As for gene E, we made the subtraction of OD 600 and seen the slope as the index which show the effects of our gene E.
Compared with gene E, killer red is more soft and safe. Based on the result, gene E tended to show property of leaking expression but killer red showed no toxin effect without either IPTG or green laser. Also, we could conclude from the data that gene E, linked with T7 promoter, restraining the growth of transformed E.coli, seemed more effective than killer red.
When it comes to gene E, efficiency of it under different promoters are tested by OD 600 which indicated the concentration of E. coli. We made subtraction of OD 600 and saw the slope as the index which show the effects of our gene E. According the slope, we could found the efficiency of Gene E under T7 promoter is between the efficiency of gene E in pBV220 induced in 37℃ and that in 42℃.
Figure 1. Efficiency of gene E in pBV220 induced in different temperatures
Figure 2. Efficiency of gene in pet28 induced by 0.5 mM IPTG
It is quite pity that we did not have enough time to compare the efficiency of gene E and killer red quantitatively, we would continue our experiment and go into more detail of these two parts in the future.
For more information about our description, please click this link: Kill_Switch
