Difference between revisions of "Part:BBa K511302"

Revision as of 02:21, 29 October 2011

Delta-mCherry Juxtacrine Signaling Ligand MammoBlock

This part encodes a fusion between the Delta-3 ligand and the monomeric mCherry red fluorescent protein.

In complex eukaryotic systems, Delta is a single-pass transmembrane protein that functions as a ligand for another single-pass receptor known as Notch. Binding of Delta to Notch results in cleavage of the intracellular domain of Notch by a ubiquitously expressed protease. The cleaved intracellular domain is then translocated to the nucleus of the Notch-bearing cell and is able to activate transcription of target genes. If Delta and Notch are expressed on the same cell, however, Delta binds to a second location on the Notch receptor, forming a complex that cannot be activated by Delta from another cell.

The C-terminal fusion of the mCherry protein to this signaling ligand means this protein is also a useful marker for transfection identification and/or membrane colocalization experiments.

Sequence and Features

Characterization

Figure 1. Activation of Notch-Gal4-ESN by Delta-mCherry in a HEK-293 Sender - CHO Receiver Co-Culture.

Figure 1 to the left shows activation of chinese hamster ovary (CHO) cells bearing Notch-Gal4-ESN driven by the CMV promoter and Citrine driven by the UAS-Gal4 promoter by human embryonic kidney (HEK-293) cells bearing Delta-mCherry driven by the Hef1a-LacO promoter. The images is an overlay of yellow fluorescence from Citrine and red fluorescence from Delta-mCherry. The abnormal morphology of red-fluorescence here was likely due to bleed-through, thus making cells appear larger than they are.