Difference between revisions of "Part:BBa K305008:Experience"

 
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<I>victorNJ</I>
 
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we (AQA_Unesp 2017) made a comparison between srfA and Pveg promoter, which is recognized as being a strong one and is already featured in the registry (BBa_K143012), and we show a relationship of cell growth and expression of GFP. As a tool for measuring the activity of promoters, reporter vectors can be required. Therefore, we chose the luciferase reporter (pBs3Clux-PsrfA), and green fluorescence protein reporter (pBs1C-PsrfA-GFP). For luciferase reporter, we did the percentage of luminescence making the ratio by the point of greatest luminescence and multiplying by 100. Already by GFP, we measured at 535nm for emission and 285nm for excitation. The readings were performed on the Infinite® M200 (Tecan)
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we (AQA_Unesp 2017) made a comparison between srfA and Pveg promoter, which is recognized  
 +
as being a strong one and is already featured in the registry (BBa_K143012),
 +
and we show a relationship of cell growth and expression of GFP.
 +
 
 +
 
 +
As a tool for measuring the activity of promoters, reporter vectors can be required. Therefore, we chose the luciferase  
 +
reporter (pBs3Clux-PsrfA), and green fluorescence protein reporter (pBs1C-PsrfA-GFP). For luciferase reporter, we did the
 +
percentage of luminescence making the ratio by the point of greatest luminescence and multiplying by 100.  
 +
Already by GFP, we measured at 535nm for emission and 285nm for excitation.  
 +
The readings were performed on the Infinite® M200 (Tecan).
 +
 
 +
 
 +
For more Details visit our [http://2017.igem.org/Team:AQA_Unesp/Parts webpage]
 +
 
 +
 
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===Characterization===
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[[Image:AQA Unesp--PvegAndSrfaOD.png|thumb|right|400px| <p align="justify"> '''Figure 1. '''''. Growth curve of Bacillus subitilis for about 7 hour, 37°C and 220 rpm.</p>]]
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[[Image:AQA Unesp--PvegAndSrfaLumi.png|thumb|right|400px| <p align="justify"> '''Figure 2. '''''. Percentage of Luminescence of PsrfA and Pveg; We use for Positive control the plasmid containing pBs3Clux without promoter. The growth has occurred for about 7 hour, 37°C and 220 rpm.</p>]]
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[[Image:AQA Unesp--SrfaODandGFP.png|thumb|right|400px| <p align="justify"> '''Figure 3. '''''. Correlation of GFP expression (pBs1C-srfA-GFP) and cell growth as a function of time for about 8 hour, 37°C and 220 rpm.</p>]]
  
  
For more Details visit our [http://2017.igem.org/Team:AQA_Unesp/Parts]
 
  
 
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Latest revision as of 10:45, 1 November 2017


Applications of BBa_K305008

User Reviews

UNIQ28beb2cc004118a8-partinfo-00000000-QINU

victorNJ

we (AQA_Unesp 2017) made a comparison between srfA and Pveg promoter, which is recognized as being a strong one and is already featured in the registry (BBa_K143012), and we show a relationship of cell growth and expression of GFP.


As a tool for measuring the activity of promoters, reporter vectors can be required. Therefore, we chose the luciferase reporter (pBs3Clux-PsrfA), and green fluorescence protein reporter (pBs1C-PsrfA-GFP). For luciferase reporter, we did the percentage of luminescence making the ratio by the point of greatest luminescence and multiplying by 100. Already by GFP, we measured at 535nm for emission and 285nm for excitation. The readings were performed on the Infinite® M200 (Tecan).


For more Details visit our [http://2017.igem.org/Team:AQA_Unesp/Parts webpage]


Characterization

Figure 1. . Growth curve of Bacillus subitilis for about 7 hour, 37°C and 220 rpm.

Figure 2. . Percentage of Luminescence of PsrfA and Pveg; We use for Positive control the plasmid containing pBs3Clux without promoter. The growth has occurred for about 7 hour, 37°C and 220 rpm.

Figure 3. . Correlation of GFP expression (pBs1C-srfA-GFP) and cell growth as a function of time for about 8 hour, 37°C and 220 rpm.


UNIQ28beb2cc004118a8-partinfo-00000002-QINU