Difference between revisions of "Part:BBa K621001:Experience"

Latest revision as of 05:25, 28 September 2011

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_K621001

Our team used this part as a glucose sensor. The part consists of an OMP-R binding region and a eYFP gene. The OMP-R binding region is regulated by the ENVZ-OMPR two component regulatory system. ENVZ, a osmolarity sensor, phosphorylates OMP-R. OMP-R then binds to the OMP-R binding region and recruits RNAP to begin transcription. However, if the osmolarity is too high the system will downregulate transcription (See part description for details). Our team found that LB media had too high of an osmolarity for testing the system. Other researchers have found this effect as well and it is well documented. The background fluorescence, when grown in LB media, was very high. To reduce background the team diluted the media by half. We found that glucose concentrations of 8% were high enough to induce downregulation and concentrations of 1% exhibited a stimulated response. Time courses revealed that a fluorescent response occurred within the first ten minutes of exposure to glucose. We also found that the most intense fluorescent response occurred at 3.5 hours in the presence of glucose. After 5 hours the cells did not seem to respond to glucose.

User Reviews

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 ===Applications of BBa_K621001===
-Our team used this part as a glucose sensor.  The part consists of an OMP-R binding region and a eYFP gene.  The OMP-R binding region is regulated by the ENVZ-OMPR two component regulatory system.  ENVZ, a osmolarity sensor, phosphorylates OMP-R.  OMP-R then binds to the OMP-R binding region and recruits RNAP to begin transcription.  However, if the osmolarity is too high the system will downregulate transcription (See part description for details).  Our team found that LB media had too high of an osmolarity for testing the system.  The background fluorescence, when grown in LB media, was very high.  To reduce background out team diluted the media in half.  Any glucose concentration higher that 2% would also begin to downregulate the system.  Time courses revealed that the most intense fluorescent response occurred at 3.5 hours in the presence of glucose.  After 5 hours the cells did not seem to respond to glucose.
+Our team used this part as a glucose sensor.  The part consists of an OMP-R binding region and a eYFP gene.  The OMP-R binding region is regulated by the ENVZ-OMPR two component regulatory system.  ENVZ, a osmolarity sensor, phosphorylates OMP-R.  OMP-R then binds to the OMP-R binding region and recruits RNAP to begin transcription.  However, if the osmolarity is too high the system will downregulate transcription (See part description for details).  Our team found that LB media had too high of an osmolarity for testing the system.  Other researchers have found this effect as well and it is well documented.  The background fluorescence, when grown in LB media, was very high.  To reduce background the team diluted the media by half.  We found that glucose concentrations of 8% were high enough to induce downregulation and concentrations of 1% exhibited a stimulated response.  Time courses revealed that a fluorescent response occurred within the first ten minutes of exposure to glucose.  We also found that the most intense fluorescent response occurred at 3.5 hours in the presence of glucose.  After 5 hours the cells did not seem to respond to glucose.
 ===User Reviews===