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BIT-China 2015

Negative Fluorescent Reporter (BBa_K1170003) Data

We mutated the EcoRI site of the promoter by single-step mutation. And we determined the activity of P-atp2 in different pH from 5.0 to 9.0 by measuring the activity of enzyme β-galactosidase(LACZ). As these bacteria grew normally and the OD₆₀₀ was up to about 1.5, we lysed cells, then added ONPG to start a color reaction and recorded the response time. After the sodium carbonate solution (1M) was added to terminate the reaction, we measured the OD₄₂₀ and OD₅₅₀, which could be put into the formula to calculate the activity(Table. 1).

Enzyme Activity= 1000*(OD₄₂₀-1.75*OD₅₅₀)/(t*0.1*OD₆₀₀)

Table. 1 The formula to calculate the enzyme activity of β-galactosidase. According to the β-galactosidase activities (Fig. 1), we can get the transcription ability of P-atp2 under the different pH environment.

Fig.1 The β-galactosidase activities in different pH from 5.0 to 9.0.

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