Date: Wed, 12 May 2004  15:19:38  \-0500
To: rd120XXXX, flybase-updatesXXXX, Annette Parks
<annettep@XXXX>
From: Kevin Cook <kcook@XXXX>
Subject: HLH106 constructs and insertions
The following information was provided by Annette Parks of Exelixis, Inc.
In P{UAS-HLH106.Exel}, wild type HLH106 (FBgn0015234) coding sequence was
cloned into P{Express-UAS}.
P{UAS-HLH106.Exel}1 is a homozygous and hemizygous viable and fertile, X
chromosome insertion.
P{UAS-HLH106.Exel}3 is a homozygous viable and fertile, third chromosome
insertion.
In P{UAS-HLH106.Ndel}, HLH106 sequences encoding a dominant negative
protein variant deleted for the highly acidic region at the N terminus
(amino acids 1-75) were cloned into P{Express-UAS}. This construct mimics a
construct of Sato et al. (J. Biol. Chem. 269: 17267, 1994).
P{UAS-HLH106.Ndel}1 is a homozygous and hemizygous viable and fertile, X
chromosome insertion.
P{UAS-HLH106.Ndel}2 is a homozygous viable and fertile, second chromosome
insertion.
P{UAS-HLH106.Ndel}3 is a homozygous viable and fertile, third chromosome
insertion.
In P{UAS-HLH106.NTdel}, HLH106 sequences were cloned into P{Express-UAS}
which encode a dominant negative protein variant deleted for the highly
acidic region at the N terminus (amino acids 1-75) and missing the
transmembrane domains.
P{UAS-HLH106.NTdel}1 is a homozygous and hemizygous viable and fertile, X
chromosome insertion.
P{UAS-HLH106.NTdel}3 is a homozygous viable and fertile, third chromosome
insertion.
In P{UAS-HLH106.Cdel}, HLH106 sequences encoding a constitutively active
protein variant truncated right before the first transmembrane domain were
cloned into P{Express-UAS}. The C terminal sequence is RGMATNSR. The
variant protein is localized to the nucleus constitutively and is
constitutively active.
P{UAS-HLH106.Cdel}1 is a homozygous and hemizygous viable and fertile, X
chromosome insertion.
P{UAS-HLH106.Cdel}2 is a homozygous viable and fertile, second chromosome
insertion.
In P{UAS-HLH106.P450}, HLH106 sequences were cloned into P{Express-UAS}
which encode a dominant negative protein variant with the rabbit cytochrome
P450 2c1 transmembrane domain (MDPVVVLGLCLSCLLLLSLWKQSYGGGKL) appended to
the C-terminus. This modification results in permanent anchoring of HLH106
protein to the endoplasmic reticulum.
P{UAS-HLH106.P450}1 is an X chromosome insertion.
P{UAS-HLH106.P450}2 is a homozygous viable and fertile, second chromosome
insertion.
P{UAS-HLH106.P450}3 is a homozygous viable and fertile, third chromosome
insertion.
__________________________________________________________
Kevin Cook, Ph.D. Bloomington Drosophila Stock Center
Department of Biology http://flystocks.bio.indiana.edu
Jordan Hall 142
Indiana University 812-856-1213
1001 E. Third St. 812-855-2577 (fax)
Bloomington, IN 47405-3700 kcook@XXXX