Subject: SCAP constructs and insertions
The following information was provided by Annette Parks of Exelixis, Inc.
In P{UAS-SCAP.Exel}, wild type SCAP (FBgn0033052) was cloned into
P{Express-UAS}. A genomic fragment was used for the C-terminal half of
SCAP; consequently, introns are still present.
P{UAS-SCAP.Exel}3 is a third chromosome insertion.
In P{UAS-SCAP.Y319C}, SCAP sequences encoding a constitutively active Y319C
variant protein were cloned into P{Express-UAS}. A genomic fragment was
used for the C-terminal half of SCAP; consequently, introns are still
present. Y319C in flies mimics hamster Y298C as described in Nohturfft et
al. (PNAS 95: 12848, 1998).
P{UAS-SCAP.Y319C}1 is a homozygous and hemizygous viable and fertile, X
chromosome insertion.
P{UAS-SCAP.Y319C}2 is a homozygous viable and fertile, second chromosome
insertion.
P{UAS-SCAP.Y319C}3 is a homozygous viable and fertile, third chromosome
insertion.
In P{UAS-SCAP.P450}, SCAP sequences encoding a dominant negative protein
variant were cloned into P{Express-UAS}. The rabbit cytochrome P450 2c1
transmembrane domain (MDPVVVLGLCLSCLLLLSLWKQSYGGGKL) was appended to the C
terminus just downstream of the final transmembrane domain to mimic a
hamster construct described by Sakai et al. (J. Biol. Chem. 10: 5785,
1998). This modification results in permanent anchoring of SCAP protein to
the endoplasmic reticulum.
P{UAS-SCAP.P450}1 is a homozygous and hemizygous viable and fertile, X
chromosome insertion.
P{UAS-SCAP.P450}3 is a homozygous viable and fertile, third chromosome
insertion.