pUASTLOTattB_VHH-GFP4::CD8::mCherry
(Plasmid #163917)

• Purpose: Expression of a membrane-tethered GFP-nanobody marked by mCherry under control of the UAS or LOP enhancers
• Depositing Lab: Markus Affolter
• Publication: Harmansa et al Nature. 2015 Nov 19;527(7578):317-22. doi: 10.1038/nature15712. Epub 2015 Nov 9.

Backbone

• Vector backbone: pUASTLOTattB
• Backbone manufacturer: Kanca, O. et.al., Raeppli: a whole-tissue labeling tool for live imaging of Drosophila development. Development (2014)
• Backbone size w/o insert (bp): 8900
• Total vector size (bp): 10700
• Vector type: Insect Expression, Cre/Lox

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Fusion of vhhGPF4 nanobody with mouse CD8 transmembrane protein and mCherry fluorophor
• Alt name: Morphotrap
• Species: M. musculus (mouse), D. melanogaster (fly)
• Tag / Fusion Protein:
  • mCherry (C terminal on insert)

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

For pUASTLOTattB_VHH-GFP4::CD8::mCherry, we inserted the VHH-GFP4 fragment after the signal peptide sequence of the mouse CD8 domain in the pUAS::CD8::GFP plasmid. We replaced the GFP by a mCherry (Clonetech) and finally cloned the VHH-GFP4::CD8::mCherry fragment into the pUASTLOTattB vector.

pUASTLOTattB vector:
Kanca, O., Caussinus, E., Denes, A. S., Percival-Smith, A. & Affolter, M. Raeppli: a
whole-tissue labeling tool for live imaging of Drosophila development.
Development 141, 472–480 (2014).

vhhGFP4 plasmid:
Saerens D, Pellis M, Loris R, Pardon E, Dumoulin M, Matagne A, Wyns L, Muyldermans S, Conrath K
J Mol Biol. 2005 Sep 23; 352(3):597-607.

mCD8 plasmid:
Lee, T. & Luo, L. Mosaic analysis with a repressible cell marker for studies of
gene function in neuronal morphogenesis. Neuron 22, 451–461 (1999).

mCherry: Clonetech