Difference between revisions of "Part:BBa K2145001:Experience"

 
 
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
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how you used this part and how it worked out.
 
  
 
===Applications of BBa_K2145001===
 
===Applications of BBa_K2145001===
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[[File:Splates95102.png]]<br>
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Qualitatively, we noticed that none of the colonies containing this plasmid showed BOTH visible RFP and visible GFP -- each colony was either red, green, or neither, but never red and green (see GG96 above). Fluorescence was observed even without the addition of inducers (IPTG and ATc), even though the reporters should be under IPTG and ATc control. We attribute this to insufficient repression by LacI and TetR.
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[[File:Alverno_ca_96.png]]
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[[File:Alverno_ca_r_96.png]]
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[[File:Alverno_ca_g_96.png]]
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Shown above are OD600 measurements, RFP fluorescence, and GFP fluorescence, respectively, for three clones containing this part in liquid culture. Again, we emphasize that only RFP OR GFP are expressed, but never both. We believe this may be due to supercoiling -- when one gene expresses, it creates supercoils that shut down the other gene.
  
 
===User Reviews===
 
===User Reviews===
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Enter the review inofrmation here.
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This part contains two fluorescent protein coding sites (for RFP and GFP) and was assembled using Golden Gate Assembly. To make plasmid, parts were taken from iGEM Registry (BBa_J04450 (RFP) and BBa_I13522 (GFP)) and assembled to make new part with a 500bp spacer in between.
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Direction of GFP: Forward, toward the spacer
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Direction of RFP: Forward, away from the spacer
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Latest revision as of 00:31, 31 October 2016


Applications of BBa_K2145001

Splates95102.png

Qualitatively, we noticed that none of the colonies containing this plasmid showed BOTH visible RFP and visible GFP -- each colony was either red, green, or neither, but never red and green (see GG96 above). Fluorescence was observed even without the addition of inducers (IPTG and ATc), even though the reporters should be under IPTG and ATc control. We attribute this to insufficient repression by LacI and TetR.

Alverno ca 96.png Alverno ca r 96.png Alverno ca g 96.png

Shown above are OD600 measurements, RFP fluorescence, and GFP fluorescence, respectively, for three clones containing this part in liquid culture. Again, we emphasize that only RFP OR GFP are expressed, but never both. We believe this may be due to supercoiling -- when one gene expresses, it creates supercoils that shut down the other gene.

User Reviews

UNIQ81cbcafc9bf3dcbe-partinfo-00000000-QINU UNIQ81cbcafc9bf3dcbe-partinfo-00000001-QINU