Contribution
We introduced the "NAN-IAV mode", an ion channel in drosophila that constitutes auditory mechanical transduction, into Saccharomyces cerevisiae BY4741, the auditory module was constructed, and Tether (SunTag scFv) was used to amplify the downstream signal to the cytoskeleton. We took the expression vector pESC-HIS in yeast as the skeleton, introduced DmNan gene module, and constructed pADH1 plasmid; The expression vector pESC-LEU in yeast was used as the skeleton to introduce DmIav and SunTag gene modules to construct pTDH3p plasmid; The expression vector pESC-URA in yeast was used as the skeleton, and the scFV and actin gene modules were introduced to construct pACT1p plasmid.

In the final digestion verification, the strip length was 12534bp by Xho I digestion, the plasmid was 3970bp and 8564bp by EcoR I digestion, and the plasmid was 7367bp and 5167bp by EcoR V digestion, which verified the correctness of pTDH3p. Linearized by Xho I digestion, the band was 9149 bp; Hind Ⅲ digestion bands were 2991 bp and 6158 bp; Sal I bands were 3031 bp and 6118 bp. The correctness of pACT1p is verified.



Fig.1pADH1 plasmid

Fig 2.pTDH3p plasmid

Fig 3.pTDH3p plasmid digestion verification

Fig.4 pACT1p plasmid

Fig.5 Validation of pACT1p plasmid digestion

Under the final fluorescence microscope, we observed that pACT1p plasmid could express sfGFP green protein in yeast, and the yeast structure was intact and grew normally, and the fusion protein was successfully expressed. The pTDH3p plasmid can express the eBFP blue protein in yeast, and the yeast structure is intact and grows normally. Most yeast structures are intact, and the fusion protein is successfully expressed.

The expression of these proteins verified our assumption of transferring drosophila gene into yeast, indicating that our experimental idea was successful.

Fig.6 Observation of Merge fluorescent protein of BY4741-ACT1 transformant

Fig7. Merge fluorescent protein observations of BY4741-ACT1 transformants

Fig8. White light DIC observation of BY4741-ACT1 transformants

Fig9. Blue fluorescence observation of BY4741 DmIav transformants

Fig10. BY4741 DmIav transformant merge fluorescent protein observations

Fig11. BWhite light DIC observation of BY4741 DmIav transformants

1. Cancer perception

Our project will focus on the expression of NAN-IAV ion channel in Saccharomyces cerevisiae. Using the "Nan IAV mode" of the ion channel that forms drosophila's auditory mechanical transduction, we will introduce Saccharomyces cerevisiae to build an auditory module to study this biological phenomenon, so as to realize mechanical stimulation of physiological activities, observe the response of yeast cells to sound waves, changes in gene level, protein level, metabolism and growth phenotype, and serve as the first round of cancer perception projects. We discussed the follow-up study of sound transduction in microorganisms. So that more patients can get more timely treatment at the early stage, and improve the treatment efficiency.

Our products are more stable than the existing electrical equipment. Electrical equipment cannot work without electricity. If we have a power failure, the equipment will fail. Our products are stable, we do not need electricity or other things, and theoretically can work for a long time.

In real life, we will use the principle that cells can make sounds, and the feature that transgenic bacteria can respond to the sounds made by cancer cells, and apply it to the instruments that detect the location of cancer cells to find cancer cells and provide help for cancer treatment. Our experiment is only the initial contribution to cancer cognition, because cancer treatment and prevention is a very important and difficult process. We hope to pave the way for future teams. We hope that future technologies or new achievements can help us transform Saccharomyces cerevisiae and provide a new method to detect cancer.

Propose the application of sound perception yeast

2. Seismic survey

Seismic wave is the spread of mechanical motion, which is generated by the elasticity of the earth medium. Its property is very close to sound wave, so it is also called ground sound wave. Earthquakes do great harm to human nature, and the self rescue time is short after the occurrence, so it is particularly important to predict earthquakes and take corresponding measures. We consider to connect this bacterium to the ground, observe seismic waves and monitor crustal activity with the help of the ground display through the transmission of electrical signals, which may be able to predict earthquake disasters and reduce the risk of erroneous measurement. At the same time, the cost of existing seismic monitors is relatively high, and the development of bacteria survey means can effectively reduce the cost.

Seismic survey