Will Deloache Test Page2: Difference between revisions
No edit summary |
|||
Line 8: | Line 8: | ||
==Specific Biological Design== | ==Specific Biological Design== | ||
[[Image:Permanent_design.png|thumb| | [[Image:Permanent_design.png|thumb|380px|right|'''Figure 1:''' Biological design of permanent memory in yeast.]] | ||
The specific biological design of this gene network is a fairly standard autoregulatory positive feedback loop. As can be seen in Figure 1 on the right, two separate plasmids were constructed that each performed separate tasks. The sensor plasmid (on the top of Figure 1) consisted of an galactose-inducible promoter (P<sub>gal</sub>) upstream of a hybrid RFP gene (Red Fluorescent Protein). Fused to the RFP gene was a DNA binding domain (DNA BD) that is specific to the P<sub>CYC1</sub> promoter, a VP64 activator region, and a nuclear localization signal (NLS). In the presence of galactose, this hybrid RFP protein would be produced and localized to the nucleus of the cell by of the NLS. Once in the nucleus, the DNA BD would allow binding of the hybrid RFP to the P<sub>CYC1</sub> promoter, at which point the VP64 activator would turn the P<sub>CYC1</sub> promoter on. | |||
The auto-feedback plasmid (on the bottom of Figure 1) consisted of a hybrid YFP gene (Yellow Fluorescent Protein) downstream of the P<sub>CYC1</sub> promoter. The same fusions (DNA BD, VP64 activator, and NLS) were made to the YFP gene that had been made the the RFP gene. Thus, the production of the hybrid YFP protein would create an autoregulatory positive feedback loop. | |||
In the presence of galactose, these yeast cells should fluorescence both red and yellow because of activation of both promoters. After galactose is removed from the cells' environment, they should retain their yellow fluorescence but lose their red fluorescence. | |||
[[Image:linebreak.png]] | |||
==Mathematical Modeling== | ==Mathematical Modeling== |
Revision as of 14:33, 4 December 2007
|