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| | ===Applications of BBa_K615000=== | | ===Applications of BBa_K615000=== |
| − | This strain is designed to test zinc finger binding using a one-hybrid selection system. It currently contains the Zif268 binding site, but that can easily be changed using lambda red or MAGE. When Zif268 fused to the omega subunit of RNA polymerase is not present, the genes His3 and URA3 are not transcribed, and thus the cells are incapable of surviving in incomplete media due to their inability to synthesize histidine. If the zinc-finger binding site promoter is leaky in some clones, URA3 can be used as a negative selector in the presence of 5-FOA: URA3 breaks down 5-FOA into a toxin, so only cells without leaky promoters will survive. Zinc finger binding can then be established by growing the strain in incomplete media and looking for a growth phenotype rescue. The sensitivity of the selection can be further manipulated by adding increasing concentrations of 3-AT, a competitive inhibitor of His3. Our results have shown that the strain is able to recognize valid binders even when diluted one in a million and in 10mM 3-AT.
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| − | [[Image: HARVhybrid1.png|thumb|left|Characterization of BBa_K61500]]
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| − | [[Image: HARVhybrid2.png|thumb|none|Sensitivity to hits among background]]
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| | ===User Reviews=== | | ===User Reviews=== |
Latest revision as of 00:57, 27 September 2011
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Applications of BBa_K615000
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