Part:BBa_K2967005:Experience

Amplifier and GFP are cloned on pcdfDuet-1 vectors which are called V-AM and V-GFP. Then empty plasmid(VE), V-GFP and V-AM are transformed into E. coli BL-21, and single colonies are added to 5 ml LB medium for overnight incubation at 37 degrees Celsius. Using 1 ml PBS to wash the bacteria and measuring the vale of OD600 to obtain cell concentration of each culture medium. The bacterial culture medium was diluted to OD600=0.025 with LBS(LB with streptomycin) and then added to black 96-well plate. IPTG with concentration of 0, 1*10-3, 1*10-4, 1*10-5 and 1*10-6 was added to induce those bacteria. After incubation for 4 hours and 6hours, the data were detected by enzyme labeling instrument. The excitation wavelength is 485 nm and the absorption wavelength is 525 nm.

Applications of BBa_K2967005

We designed transcriptional amplifier (Amp) employed to amplify the transduced transcriptional signal from the NO sensors or any other biosignals. Adding the Amp to the downsream of the specific sensor genes, the input will be amplified and the output or gene products will be accelerated.

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