Part:BBa_K1510233:Experience

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Applications of BBa_K1510233

Six plates of biosafety E. coli was cultured in individual 10 microliter LB. Two in a group one test one control. Therefore, we have three groups of data (6 Control / 6 Test ) , (12 Control / 12 Test ) , (24 Control / 24 Test ) All six of them are covered with Aluminum foil, in order to isolate blue light disturbance.We culture our E. coli in an incubator with shaker and light bulb to provide favored environment.

After the culture reaches 6, 12 and 24 hour, we open the aluminum foil of the test group and plotted the surviving cell number over blue light exposure time. The counting procedure was carried out by CFU (colony Forming Unit), and te surviving ratio was documented over light exposure time.

And we did serial dilution for every sample we acquired. And apply our 10^4, 10^5, 10^6, 10^7, 10^8, 10^9, 10^10 dilutioned liquid culture to the plate.

he overall results “group data 24 Hour” and the colony counting list was placed in the down-right.

Results

The Clock Count Dilution Based test /// CCDB test Due to the killing efficiency attained less than 1 survival in every 100 population at the growing time 6 hour. The CFU counting may not be a suitable alternative. Therefore, we did rough estimation by a serial 10X times dilution. By setting the gradient at 10 times dilution, we can know where the cell number’s power index lye. For example, the colony disappear between gradient of 4 and 5, thus we can infer the cell number X would be : 104 < X < 105 . So on and on, there are several examples in graph.

Inferred from the above table, we could discover that after exposure to light for a period of time, the population of modified E. coli(transformed by ccdb suicide gene) decrease. The more exposure time, the less the population is. So we could conclude that our modified E. coli could be killed by exposure to light.Results

Indicated from the above graph, we could discover that after exposure to light for a period of time, the population of E. coli(transformed by ccdb suicide gene) dramatically decrease. And even the population of E. coli reached the stable stage, we can still attain 92.5%proportion of killing.

Inferred from the above graph, we could discover that after exposure to light for a period of time, the survival rate of E. coli(transformed by ccdb suicide gene) dramatically decrease. And the less culture time, less time is needed to kill most of them (In 2hr survival rates are all below 1%). So we could conclude that our modified E. coli could be easily killed by exposure to light in a short upon released.

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