Team:SIAT-SCIE/Proof

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Overview

Our project is mainly focusing on:

• The ability of CAHS proteins in increasing the survival rate of desiccated bacteria
• The ability of Dsup proteins in increasing the survival rate of irradiated bacteria
• The amount of Dsup proteins in affecting the survival rate of irradiated bacteria

Experiment & Results

Desiccation Tolerance Experiment

After the vector construction and protein expression, we dilute the broth and spread it evenly to the growth culture, to count the Clone Forming Unit per 100μl of diluted broth. And use the same tube of broth, dehydrate it by centrifuge and desiccator. After leaving the dried broth for 48 hours in room temperature, we rehydrate it and spread it evenly to the growth culture again, and we can obtain the Clone Forming Unit per 100μl of diluted desiccated broth. By these two data, we can calculate the survival rate of the bacteria after desiccation.

Results:

Optical Density of Broth

Clone formed before desiccation

Clone formed after desiccation

Radiation Tolerance Experiment

The expression is induced by different concentration of IPTG, and use a microplate reader to obtain the fluorescence, which can indicate the amount of protein expressed. Then we dilute the broth, take 100μl of diluted broth, spread them on the growth culture, to obtain the original Clone Forming Unit per 100μl diluted broth. Then take 100μl of diluted broth, spread them on the growth culture. Place the growth culture under UV for different time periods, then grow over night, to obtain the Clone Forming Unit per 100μl of diluted irradiated broth. From these experiments, we can count the survival rates of the bacteria with different amount of Dsup protein expressed and survival rates of bacteria after different amount of radiation irradiated.

Results:

OD:

Fluorescence

Clone formed before irradiation

Clone formed after irradiation

Conclusion:

Now, we are still doing our experiment, so it is hard to say whether our theory works. Let us pray that the paper is true and we did not make any mistake.