Author: Prashali Bansal; Johannes Madlung; Kristina Schaaf; Boris Macek; Fulvia Bono
Title: An interaction network of RNA-binding proteins involved in Drosophila oogenesis Document date: 2020_1_9
ID: 2f9nc2to_42
Snippet: Typically, we co-expressed an EGFP (referred to as GFP)-tagged bait with an HA-tagged candidate protein and used the GFP-TRAP system (Chromotek) for IPs. We observed that small proteins (<25kDa) expressed poorly as fusions with HA or HA-Flag. Substitution with the GFP tag improved the expression in all the cases tested. To be able to validate the interactions of such small proteins, we co-expressed an HA-Flag-tagged candidate with a GFP-tagged ba.....
Document: Typically, we co-expressed an EGFP (referred to as GFP)-tagged bait with an HA-tagged candidate protein and used the GFP-TRAP system (Chromotek) for IPs. We observed that small proteins (<25kDa) expressed poorly as fusions with HA or HA-Flag. Substitution with the GFP tag improved the expression in all the cases tested. To be able to validate the interactions of such small proteins, we co-expressed an HA-Flag-tagged candidate with a GFP-tagged bait and performed the IP with anti-Flag (Fig. 7f, h) . Since Vas does not express well as an HA-or HA-Flag fusion, the small proteins could not be assayed efficiently for interaction with Vas.
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