What are Gal4 GFP enhancer trap lines?
These two enhancer trap lines are very useful tools with which to study the molecular and cellular bases of lateral root development using targeted gene expression. These lines were used for genetic ablation experiments by targeting the expression of a toxin-encoding gene.
How does Gal4 UAS system work?
The Gal4/UAS system Gal4 is a transcriptional activator that binds to UAS enhancer sequences found in DNA. It then recruits transcription machinery to the site to induce gene expression. Thus, genes and siRNA encoded downstream of the UAS sequence are only expressed when Gal4 is expressed.
What are enhancer trap lines?
Why is Gal4 UAS used?
GAL4 and the UAS are very useful for studying gene expression in Drosophila as they are not normally present and their expression does not interfere with other processes in the cell.
What is the GAL4?
The cell body can be visualised using tyrosine hydroxylase (TH) antibody -an enzyme involved in the synthesis of dopamine -and ddc-gal4 and TH-Gal4, which are dopaminergic neurone-specific Gal4 drivers. Studies of neurodegenerative diseases using Drosophila and the development of novel approaches for their analysis.
What does GAL4 bind to?
The GAL4 protein of yeast activates the transcription of several genes involved in galactose metabolism. This event requires that GAL4 bind to upstream activation sites with the consensus sequence 5′-CGGN5(T/A)N5CCG-3′.
What genes does GAL4 regulate?
The Gal4 transcription factor is a positive regulator of gene expression of galactose-induced genes. This protein represents a large fungal family of transcription factors, Gal4 family, which includes over 50 members in the yeast Saccharomyces cerevisiae e.g. Oaf1, Pip2, Pdr1, Pdr3, Leu3.
What is the role of Gal4?
Gal4 activates transcription by recruiting coactivators and the general transcription machinery to promoter regions through its activation domain (Fig 1). Important questions remain as to what are the functional targets of Gal4 and whether it contacts one or many proteins during the process of recruitment.
What are enhancer traps used for?
An enhancer trap is a method in molecular biology. The enhancer trap construct contains a transposable element and a reporter gene. The first is necessary for (random) insertion in the genome, the latter is necessary for identification of the spatial regulation by the enhancer.
What is beta geo?
The β-GEO marker is a fusion protein encoded by lacZ and the neomycin resistance gene. To prevent the secretion of the selection marker when integrated into a secretory pathway gene, a transmembrane domain is added to the trap vector anchoring the secretory (marker) protein to the cell membrane [89].
What is the role of GAL4?
How many GAL4 GFP enhancer trap lines are there?
To facilitate the monitoring of guard cells during development and isolation, a population of 704 GAL4 GFP enhancer trap lines was screened and four single insert lines with guard cell GFP expression and one with developmentally-regulated guard cell GFP expression were identified.
What is the role of gal4-gfp enhancer in root development?
GAL4-GFP enhancer trap lines for genetic manipulation of lateral root development in Arabidopsis thaliana Lateral root development occurs throughout the life of the plant and is responsible for the plasticity of the root system.
What is a gal4-activated promoter?
A chosen target gene can be placed under the control of a GAL4-activated promoter ( U pstream A ctivation S equences, UAS), transformed into Drosophila and maintained silently in the absence of GAL4. Genetic crosses between this single line and an enhancer trap line specifically activates the target gene in a chosen tissue or cell type.
Can GAL4 be used for targeted gene expression?
Targeted gene expression using GAL4 is a powerful tool to analyse the molecular and cellular bases of development. However, it depends on the isolation of suitable GAL4 expressing lines (enhancer trap or promoter).