Difference between revisions of "Part:BBa K2997012"

(→‎Characterization)
Line 5: Line 5:
 
===Characterization===
 
===Characterization===
  
−
This year, we characterized FNR promoter (BBa_K1123000) which is an anaerobic promoter. Originally, we used this promoter to drive the expression of TAL constructs. However, finding that this promoter has a higher expression level of GFP under aerobic conditions, we did not use this promoter for our project. Instead, we used the strong constitutive promoter J23100 as our promoter.
+
This year, we characterized FNR promoter ([[Part: BBa_K1123000|BBa_K1123000]]) which is an anaerobic promoter. Originally, we used this promoter to drive the expression of TAL constructs. However, finding that this promoter has a higher expression level of GFP under aerobic conditions, we did not use this promoter for our project. Instead, we used the strong constitutive promoter J23100 as our promoter.
  
−
We first cultured DH5a pSB1C3-Pfnr-GFP under aerobic and anaerobic conditions. After 10 hours of incubation, we fixed the E. coli cells in 2% agarose gel and placed it on a glass slide, then place the glass slide under a microscope for image capturing.  
+
We first cultured DH5a pSB1C3-P<sub>FNR</sub>-GFP under aerobic and anaerobic conditions. After 10 hours of incubation, we fixed the <i>E. coli</i> cells in 2% agarose gel and placed it on a glass slide, then place the glass slide under a microscope for image capturing.  
  
 
<html>
 
<html>
Line 16: Line 16:
 
<br>
 
<br>
 
</html>
 
</html>
−
Fig. 1 Differential interference contrast (DIC) microscope image and fluorescence microscope image for a single E. coli cell.
+
Fig. 1 Differential interference contrast (DIC) microscope image and fluorescence microscope image for a single <i>E. coli</i> cell.
  
−
We then compared the brightness of a single E. coli cell under a fluorescence microscope using ImageJ Intensity Processing. The results are shown below.
+
We then compared the brightness of a single <i>E. coli</i> cell under a fluorescence microscope using ImageJ Intensity Processing. The results are shown below.
  
 
<html>
 
<html>
Line 27: Line 27:
 
<br>
 
<br>
 
</html>
 
</html>
−
Fig. 2. Quantification of single E. coli cell (n=3) fluorescence intensity using ImageJ Intensity Processing after 10 hours of incubation in both aerobic and anaerobic conditions.  
+
Fig. 2. Quantification of single <i>E. coli</i> cell (n=3) fluorescence intensity using ImageJ Intensity Processing after 10 hours of incubation in both aerobic and anaerobic conditions.  
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 09:07, 20 October 2019


Pfnr-GFP

Characterization

This year, we characterized FNR promoter (BBa_K1123000) which is an anaerobic promoter. Originally, we used this promoter to drive the expression of TAL constructs. However, finding that this promoter has a higher expression level of GFP under aerobic conditions, we did not use this promoter for our project. Instead, we used the strong constitutive promoter J23100 as our promoter.

We first cultured DH5a pSB1C3-PFNR-GFP under aerobic and anaerobic conditions. After 10 hours of incubation, we fixed the E. coli cells in 2% agarose gel and placed it on a glass slide, then place the glass slide under a microscope for image capturing.



Fig. 1 Differential interference contrast (DIC) microscope image and fluorescence microscope image for a single E. coli cell.

We then compared the brightness of a single E. coli cell under a fluorescence microscope using ImageJ Intensity Processing. The results are shown below.



Fig. 2. Quantification of single E. coli cell (n=3) fluorescence intensity using ImageJ Intensity Processing after 10 hours of incubation in both aerobic and anaerobic conditions.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 39
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 790