Difference between revisions of "Part:BBa K1363400"
Line 2: Line 2:
 
<partinfo>BBa_K1363400 short</partinfo>
 
<partinfo>BBa_K1363400 short</partinfo>
  
 +
<html>
 
<p>This part is our integral blue light pathway. It consists of a blue light inducing amilCP expression system, a blue light sensor where YF1 and FixJ sense the light and lambda CI acts as the not gate to control blue protein expression.</p>
 
<p>This part is our integral blue light pathway. It consists of a blue light inducing amilCP expression system, a blue light sensor where YF1 and FixJ sense the light and lambda CI acts as the not gate to control blue protein expression.</p>
 
<p>The circuit is as following: We combine the light sensor and amiICP expression system on the same plasmid and it can let the bacteria complete blue photography independently.  </p>
 
<p>The circuit is as following: We combine the light sensor and amiICP expression system on the same plasmid and it can let the bacteria complete blue photography independently.  </p>
Line 27: Line 28:
 
<p>We may conclude that neither red nor green light has repressive effects on blue inducible bacteria</p>
 
<p>We may conclude that neither red nor green light has repressive effects on blue inducible bacteria</p>
 
https://static.igem.org/mediawiki/parts/d/de/USTC-Grow.png"
 
https://static.igem.org/mediawiki/parts/d/de/USTC-Grow.png"
 +
</html>
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===

Revision as of 05:17, 1 November 2014

R0010-K592016-B0015-K502020

This part is our integral blue light pathway. It consists of a blue light inducing amilCP expression system, a blue light sensor where YF1 and FixJ sense the light and lambda CI acts as the not gate to control blue protein expression.

The circuit is as following: We combine the light sensor and amiICP expression system on the same plasmid and it can let the bacteria complete blue photography independently.

https://static.igem.org/mediawiki/parts/5/50/USTC-5.jpg

In the blue light-inducible system, FixK2 promoter is the downstream signaling promoter of YF1/FixJ blue light-sensing system. The YF1/FixJ system becomes inactive when illuminated by blue otherwise active. An inverter is placed behind FixK2 promoter. 
In the absence of blue light, YF1 phosphorylates FixJ that activates FixK2 promoter, allowing the transcription of lambda CI, thus repressing the amilCP output (Blue Chromophore Protein).

Experiments:

1. Test in solid media

Without induced expression, bacteria containing blue light sensing-imaging system grew up and developed in the LB media. Then we divided the media into two regions, one of them received blue light simulation while the other one kept darkness by a base plate to block blue light. The testing conditions are showed in the following

https://static.igem.org/mediawiki/parts/3/3d/USTC-2.png

After 12-hours' development, the result showed the bacteria could produce blue chromo-protein when stimulated by blue light and kept its color in the darkness:

https://static.igem.org/mediawiki/parts/9/95/USTC-3.png

2. Test in liquid media

The idea of control experiment is the same as that ain solid media. We prepared two test tubes containing equivalent bacterium solution. The test was conducted at 18:00, August 4th when one of them was put under blue light and the other was developed in the darkness.

At 11:00, August 5th, the tube stimulated under blue light turned light blue while the other one kept the original color.

After 24 hours, the blue bacteria showed evidently. Using centrifuge to get the bacteria showed extremely distinct blue chromo-protein expression based on blue light activation was come true.

https://static.igem.org/mediawiki/parts/9/94/USTC-1.png

3. Pattern Test

Test of a blue sun pattern focused on sharp shape of a picture is in the fo llowing. The result showed bacteria could produce protein at the edge of the picture accurately.

https://static.igem.org/mediawiki/parts/c/c1/USTC-4.jpg

4.The interference of other color

In order to see if other colors will influence the blue inducible bacteria, we observed cell populations containing the blue light inducible system grown in four tubes under same conditions.However are differently illuminated with red light, green light and blue light and in dark. Their growing curves are approximately the same with acceptable deviation. The growth of dark exposed bacteria slightly surpassed others since they eat a lot and produce less, which should has nothing to do with repress from the light.

We may conclude that neither red nor green light has repressive effects on blue inducible bacteria

https://static.igem.org/mediawiki/parts/d/de/USTC-Grow.png" Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 770
    Illegal NgoMIV site found at 842
    Illegal NgoMIV site found at 932
    Illegal NgoMIV site found at 950
    Illegal NgoMIV site found at 1462
    Illegal NgoMIV site found at 1755
    Illegal NgoMIV site found at 1849
    Illegal AgeI site found at 484
    Illegal AgeI site found at 1630
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1519
    Illegal BsaI.rc site found at 383