Drosophila Proteome Project

1
Drosophila Proteome Project Harvard,
Berkeley, Cellzome (Heidelberg), TATA Institute

• Goal Define protein complex map
• Tag a protein (cDNA), express, affinity
  purify protein and associated partners,
• define protein complex elements

S Artavanis Tsakonas
2
Aims

• TAP tag about 1/3 of Dm genes (Gold set) in Mt
• and UAS vectors both at the 5 and 3 ends
• Establish stably transfected lines in KC cell
• Affinity purification of complexes from
  transgenic
• KC cell lines
• Analysis of complexes by Mass spectrometry
• Generate corresponding transgenic fly lines
• Generate protein complex

S Artavanis Tsakonas
3
Working premises

• With 1/3 tagged proteins we will see the entire
  proteome
• Analysis will be first done with only c
  terminally tagged entry points. Based on
  previous experience we expect that 20 of those
  proteins will be inactive. We expect to refine
  analysis as necessary (N terminally tagged
  proteins will be available)
• Establish base line with KC cells. Generate
  transgenic flies (UAS) for more in depth analysis
  but such analysis is not part of the initial
  project.
• Everything available to the community as soon
  (explain) as produced reagents and information.
  Every 3 months release. No intellectual property
  no strings attached.
• We aim at 4 years

S Artavanis Tsakonas
4
Main Participants

• Spyros Artavanis-Tsakonas
• Sue Celniker
• Mark Stapleton
• Georg Casari
• Anne Claude Gavin
• Bernhard Kuester
• Vijay Raghavan
• Gerry Rubin

S Artavanis Tsakonas