This chapter focuses on one particular approach, the gene trap screen in mouse embryonic stem (ES) cells, for discovering genes that are specifically expressed during vascular development.
The transgenic Ubx-YFP (CPTI-000601) and Hth-YFP (CPTI-000378) FlyProt protein trap lines were generated via a transposon-based exon-trapping screen [30]; details of these lines are available from http://www.flyprot.org/.org/
In a large scale enhancer trap screen, 6966 insertions were analyzed by iPCR.
EtMn16 is a product of a Sleeping Beauty (SB) transposon mediated-enhancer trap screen previously described [6].
The notion was to attempt to enrich for possible mutants through a fat body insertional enhancer trap screen followed by a comprehensive longevity screen.
Promoterless DsRed was amplified from pDsRed Monomer (Clontech, Mountain View, CA) and cloned into pBR322 for use in the promoter trap screen.
First, we performed a two-component (minimal promoter Gal4; UAS-GFP) F1 enhancer trap screen [31]; isolating lines with GFP expression in the fat body (Figure 1A).
To identify regulators of lifespan expressed in sites that regulate metabolism, we designed a multi-tiered approach, in which the initial step was a minimal promoter-Gal4; UAS-GFP fat body enhancer trap screen [31].
This gene was isolated in a gene trap screen for novel transmembrane proteins (mouse line KST37) and was of particular interest based on its protein architecture and expression pattern [8], [9].
This genomic region was not predicted by the Berkeley Drosophila Genome Project (R5.19) to contain a mdg3 natural transposon, and hence represents a divergence between the sequenced strain and the strain used for the GAL4 enhancer trap screen.
In a D. melanogaster fat body insertional mutagenic enhancer trap screen designed to isolate genes that control longevity, we identified a regulator of G protein signaling (RGS) domain containing sorting nexin, termed snazarus (sorting nexin lazarus, snz).
