The P{UAS-LT3-Dam-RpII215} construct is used in the "Targeted DamID" ("TaDa") technique, in which translation of the Ecol\dam protein at very low levels in a cell- or tissue-specific fashion enables genome-wide profiling of DNA- or chromatin-binding proteins without requiring cell sorting, fixation or affinity purification and avoiding the toxicity associated with higher levels of Ecol\dam expression. In the TaDa system, a primary open reading frame (ORF1) is followed by two stop codons and a secondary ORF (ORF2) which includes Ecol\dam fused to a DNA- or chromatin-binding protein of interest. The presence of ORF1 greatly reduces the translation of the Ecol\dam-containing ORF2. In the P{UAS-LT3-Dam-RpII215} construct, the first ORF encodes full-length mCherry, the second ORF is a fusion protein consisting of the RpII215 gene fused to the Ecol\dam ORF and tagged with Tag:MYC, and the regulatory sequences that drive expression are UAS sequences.
The P{UAS-LT3-Dam-RpII215} construct is used in the "Targeted DamID" ("TaDa") technique, in which translation of the Ecol\dam protein at very low levels in a cell- or tissue-specific fashion enables genome-wide profiling of DNA- or chromatin-binding proteins without requiring cell sorting, fixation or affinity purification and avoiding the toxicity associated with higher levels of Ecol\dam expression. In the TaDa system, a primary open reading frame (ORF1) is followed by two stop codons and a secondary ORF (ORF2) which includes Ecol\dam fused to a DNA- or chromatin-binding protein of interest. The presence of ORF1 greatly reduces the translation of the Ecol\dam-containing ORF2. In the P{UAS-LT3-Dam-RpII215} construct, the first ORF encodes the full-length mCherry form of Disc\RFP, the second ORF is a fusion protein consisting of the RpII215 gene fused to the Ecol\dam ORF and tagged with T:Hsap\MYC, and the regulatory sequences that drive expression are Scer\UAS sequences.