Generator

Part:BBa_K4202045:Experience

Designed by: Zhijian Yan, Zhuoyun Ren   Group: iGEM22_ZJU-China   (2022-10-03)
Revision as of 13:05, 12 October 2022 by Simplest (Talk | contribs) (→‎Method)


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Method

Measurement device construction

The sequence of this part was synthesis by Genscript and constructed to the pHY300PLK by In-Fusion clone.

Characterization

The transformant (Bacillus subtilis with pHY300PLK empty plasmid and kill switch) were inoculate to 2ml LB medium and cultured in the test tube overnight. The next day, the culture were inoculated into 2ml TB medium containing 1% sucrose (w/v) at the ratio of 1:100. After the OD600nm(L=1mm) reaches around 0.1, the OD600nm(L=1mm) were measured every 40minutes.

Result

Compared to the control group, the OD600nm(L=1mm) of strains containing the kill switch began to decrease after the 600nm(L=1mm) reached 0.12. The early decrease of OD600nm indicated that the bacteria are dying, which means our kill switch is working.

Kill result.png
Fig 1The growth curve of two Bacillus subtilis WB600 strains.

Blue curve represents strains containing suicide switches and the yellow line represents strains containing the empty plasmid(CK)


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