Difference between revisions of "Part:BBa K1170003:Experience"
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== '''Team BIT-China 2015 ''' ==
 
== '''Team BIT-China 2015 ''' ==
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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 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).
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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) was added 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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Enzyme Activity= 1000*(OD<sub>420</sub>-1.75*OD<sub>550</sub>)/(t*0.1*OD<sub>600</sub>)
 
Enzyme Activity= 1000*(OD<sub>420</sub>-1.75*OD<sub>550</sub>)/(t*0.1*OD<sub>600</sub>)
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Table. 1 The formula to calculate the enzyme activity of β-galactosidase.
 
Table. 1 The formula to calculate the enzyme activity of β-galactosidase.
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According to the β-galactosidase activities (Fig. 1), we can get the transcription ability of P-atp2 under the different pH environment.
 
According to the β-galactosidase activities (Fig. 1), we can get the transcription ability of P-atp2 under the different pH environment.
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https://static.igem.org/mediawiki/parts/9/9c/BIT_China_parts_P-atp2.png
 
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Fig.1 The β-galactosidase activities in different pH from 5.0 to 9.0.
 
Fig.1 The β-galactosidase activities in different pH from 5.0 to 9.0.
  
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Team BIT-China 2015 improved this part by making it compatible with RFC10 (see: BBa_K1675022)
 
  
  
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Revision as of 15:12, 17 September 2015

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K1170003

User Reviews

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Team BIT-China 2015

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.


BIT_China_parts_P-atp2.png

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



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