Difference between revisions of "Part:BBa K1170003:Experience"
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| − | + | The verification of alkali-induced promoter P-atp2's transcription function (<partinfo>BBa_K1170003</partinfo>) Data (BIT-China) | |
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| − | 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 OD<sub>600</sub> 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) | + | We mutated the <i>EcoRI</i> 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 OD<sub>600</sub> was up to about 1.5, we lysed cells, then added ONPG to start a color reaction and recorded the response time. After adding the sodium carbonate solution (1M) to terminate the reaction, we measured the OD<sub>420</sub> and OD<sub>550</sub>, which could be put into the formula to calculate the activity(Table. 1). |
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Latest revision as of 19:41, 18 September 2015
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BIT-China 2015 |
The verification of alkali-induced promoter P-atp2's transcription function (BBa_K1170003) Data (BIT-China)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 adding the sodium carbonate solution (1M) 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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