Figure 1. The Drosophila Protein interaction Map (DPiM) project’s protein complex purification pipeline. All the sequence verified Drosophila Gold Collection cDNA clones were transferred to a pDNR-Dual vector that allows donor clones to be transferred via Cre-lox recombination into acceptor vectors as distinct C-terminal or N-terminal tagged expression constructs. A collection of different acceptor vectors has been created with choice of affinity tags (FLAG-HA tag, TAP-tag, His-tag etc.) to be used as expression clones in cell culture (as metallothionein-inducible constructs) or to generate transgenic flies (as UAS expression constructs). Nearly 5,000 individual FLAG-HA tagged proteins were expressed in S2R+ cells followed by co-affinity purification and mass spectrometry to identity protein partners and construction of DPiM. The same set of clones has been used to generate a complementary set of stable transgenic lines for tissue and stage specific expression using the UAS-Gal4 system.