Subject: lacZ constructs and insertions
The following information was provided by Annette Parks of Exelixis, Inc.
In P{ey3.5-lacZ}, lacZ was cloned into EcoR1/Bgll2 sites in the polylinker
of P{Express-ey3.5}. The lacZ gene was isolated in two fragments:
EcoR1-Sac1 was from pC4-AUG-betagal (FBmc0000220) and is an in-frame fusion
of the Adh leader to the 5' end of lacZ. The other half of lacZ was from
pDCZA (a clone from Dan Curtis) and has the 3' end of lacZ directly fused
to the Adh 3'UTR immediately following the stop codon.
P{ey3.5-lacZ}2 is a homozygous viable and fertile, second chromosome insertion.
P{ey3.5-lacZ}3 is a homozygous viable and fertile, third chromosome insertion.
In P{vgM-lacZ.Exel}, lacZ was cloned into EcoR1/Bgll2 sites in the
polylinker of P{Express-vgM}. The lacZ gene was isolated in two fragments:
EcoR1-Sac1 was from pC4-AUG-betagal (FBmc0000220) and is an in-frame fusion
of the ADH leader to the 5' end of lacZ. The other half of lacZ was from
pDCZA (a clone from Dan Curtis) and has the 3' end of lacZ directly fused
to the ADH 3'UTR immediately following the stop codon.
P{vgM-lacZ.Exel}2 is a second chromosome insertion.
P{vgM-lacZ.Exel}3 is a homozygous viable and fertile, third chromosome
insertion.
In P{GMR-UAS-lacZ}, a UAS-Hsp70TATA-lacZ fragment was cloned into
P{Express-glass} in place of the Hsp70 TATA sequence of that vector using
KpnI and SacII. The result was GMR-UAS-Hsp70TATA-lacZ. Both UAS and GMR
can drive lacZ expression.
P{GMR-UAS-lacZ}2 is a second chromosome insertion.
P{GMR-UAS-lacZ}3 is a homozygous viable and fertile, third chromosome
insertion.
In P{Dll-UAS-lacZ}, a Distalless (FBgn0000157) enhancer was placed 5' of
UAS-Hsp70TATA-lacZ in P{Express} using XhoI and KpnI. lacZ is expressed in
the embryonic Dll pattern, but not in imaginal disks. UAS can also drive
lacZ expression.
P{Dll-UAS-lacZ}1 is a homozygous and hemizygous viable and fertile, X
chromosome insertion.
In P{dpp-lacZ.Exel.1} and P{dpp-lacZ.Exel.2}, the dpp disk enhancer
described in St. Johnston et al., 1990 (Genes Dev. 4: 1114-1127;
FBrf0051824) drives lacZ expression. In P{dpp-lacZ.Exel.1}, the relative
orientation of the 3 kb enhancer fragment to lacZ matches the orientation
of the enhancer and the dpp coding region in vivo and in P{TnJMB7}
(FBtp0004773) described in Masucci et al., 1990 (Genes. Dev. 4: 2011-2023;
FBrf0051842). In P{dpp-lacZ.Exel.2}, the enhancer is in the opposite
orientation and matches that in P{TnJMB2} (FBtp0004772) also described in
Masucci et al.
P{dpp-lacZ.Exel.1}2 is a homozygous viable and fertile, second chromosome
insertion.
P{dpp-lacZ.Exel.2}1is an X chromosome insertion.
P{dpp-lacZ.Exel.2}2 is a homozygous viable and fertile, second chromosome
insertion.
P{dpp-lacZ.Exel.2}3 is a homozygous viable and fertile, third chromosome
insertion.