Barrett constructs and insertions. - Barrett, K. (2005)
The following information accompanied stocks donated to the Bloomington Stock Center by Kathy Barrett, University College London (11/05). PUASp-T7.RhoGEF2 expresses RhoGEF2 protein with an N-terminal T7 tag. The construct was cloned in PUASp. P{UASp-T7.RhoGEF2}5 is a homozygous viable and fertile, third chromosome insertion. P{UASp-T7.RhoGEF2}6b is a homozygous viable and fertile, second chromosome insertion. PUASp-T7.RhoGEF2.DeltaPDZ expresses RhoGEF2 protein lacking PDZ domain (amino acids 256-335) with an N-terminal T7 tag. The construct was cloned in PUASp. P{UASp-T7.RhoGEF2.DeltaPDZ}4a is a homozygous viable and fertile, third chromosome insertion. PUASp-FLAG.Rho1 expresses Rho1 protein with an N-terminal FLAG tag. The construct was cloned in PUASp. P{UASp-FLAG.Rho1}2a is a second chromosome insertion. P{UASp-FLAG.Rho1}5 is a homozygous viable and fertile, third chromosome insertion. PUASp-FLAG.Rho1.D100N expresses Rho1 protein with a D100N amino acid change and an N-terminal FLAG tag. The construct was cloned in PUASp. The D100N amino acid change generates a temperature-sensitive Rho1 protein. This mutation mimics a temperature-sensitive RHO1 mutation in Saccharomyces cerevisiae described by Ozaki et al. ("Rom1p and Rom2p are GDP/GTP exchange proteins (GEPs) for the Rho1p small GTP binding protein in Saccharomyces cerevisiae", EMBO J. 1996; 15:2196-207). P{UASp-FLAG.Rho1.D100N}3 is a homozygous viable and fertile, third chromosome insertion. PUASp-GFP.Rho1 expresses Rho1 with an N-terminal GFP tag. The construct was cloned in PUASp. Fluorescence is detectable in wing imaginal discs 30 hours after puparium formation. P{UASp-GFP.Rho1}1 is a homozygous and hemizygous viable and fertile, X chromosome insertion. P{UASp-GFP.Rho1}6 is a homozygous viable and fertile, third chromosome insertion.