Difference between revisions of "Part:BBa K511302"
(rollbackEdits.php mass rollback) |
|||
| Line 1: | Line 1: | ||
| − | + | __NOTOC__ | |
| + | <partinfo>BBa_K511302 short</partinfo> | ||
| + | |||
| + | 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. | ||
| + | |||
| + | <!-- Add more about the biology of this part here | ||
| + | ===Usage and Biology=== | ||
| + | |||
| + | <!-- --> | ||
| + | <span class='h3bb'>Sequence and Features</span> | ||
| + | <partinfo>BBa_K511302 SequenceAndFeatures</partinfo> | ||
| + | |||
| + | |||
| + | <!-- Uncomment this to enable Functional Parameter display | ||
| + | ===Functional Parameters=== | ||
| + | <partinfo>BBa_K511302 parameters</partinfo> | ||
| + | <!-- --> | ||
| + | |||
| + | ==Characterization== | ||
| + | [[Image:NotchDeltaActivation.png|thumb|left|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. | ||
| + | |||
| + | <html><br><br><br><br><br><br><br><br><br><br> | ||
| + | |||
| + | <center><b>Timelapse of CHO cells transfected with CMV:tTA and TRE:Delta-mCherry with dox<br> | ||
| + | <iframe width="420" height="315" src="http://www.youtube.com/embed/oHJhDiTZZCI" frameborder="0" allowfullscreen></iframe></center> | ||
| + | </b><br> | ||
| + | The video is a timelapse of CHO cells transfected with CMV:tTA and TRE:Delta-mCherry. At t=0, 100ug/mL dox is added to the media. The movie is a timelapse sped up such that one second of movie time is one hour of real time. | ||
| + | |||
| + | </html> | ||
Latest revision as of 16:18, 10 May 2013
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
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 85
Illegal PstI site found at 162
Illegal PstI site found at 1005
Illegal PstI site found at 1428
Illegal PstI site found at 2105
Illegal PstI site found at 2491 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 85
Illegal PstI site found at 162
Illegal PstI site found at 1005
Illegal PstI site found at 1428
Illegal PstI site found at 2105
Illegal PstI site found at 2491
Illegal NotI site found at 494 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 85
Illegal PstI site found at 162
Illegal PstI site found at 1005
Illegal PstI site found at 1428
Illegal PstI site found at 2105
Illegal PstI site found at 2491 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 85
Illegal PstI site found at 162
Illegal PstI site found at 1005
Illegal PstI site found at 1428
Illegal PstI site found at 2105
Illegal PstI site found at 2491
Illegal AgeI site found at 521 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 33
Illegal BsaI site found at 473
Illegal BsaI.rc site found at 1738
Illegal BsaI.rc site found at 1910
Illegal SapI site found at 1226
Illegal SapI.rc site found at 1998
Characterization
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.
The video is a timelapse of CHO cells transfected with CMV:tTA and TRE:Delta-mCherry. At t=0, 100ug/mL dox is added to the media. The movie is a timelapse sped up such that one second of movie time is one hour of real time.