Generation of Driver and Reporter Constructs for the GAL4 Expression System in Drosophila

2008 ◽  
Vol 2008 (8) ◽  
pp. pdb.prot5029-pdb.prot5029 ◽  
Author(s):  
T. D. Southall ◽  
A. H. Brand
Keyword(s):  
Expression System ◽  
Gal4 Expression

scholarly journals Vitellogenin family gene expression does not increase Drosophila lifespan or fecundity

F1000Research ◽  
2014 ◽  
Vol 3 ◽  
pp. 125 ◽  
Author(s):  
Yingxue Ren ◽  
Kimberly A. Hughes
Keyword(s):  
Meta Analysis ◽  
Expression System ◽  
High Fecundity ◽  
Protein Precursor ◽  
Aging Studies ◽  
Competing Demands ◽  
Extended Lifespan ◽  
Gal4 Expression ◽  
Queen Bees ◽  
Family Gene

One of the most striking patterns in comparative biology is the negative correlation between lifespan and fecundity observed in comparisons among species. This pattern is consistent with the idea that organisms need to allocate a fixed energy budget among competing demands of growth, development, reproduction and somatic maintenance. However, exceptions to this pattern have been observed in many social insects, including ants, bees, and termites.  In honey bees (Apis mellifera), Vitellogenin (Vg), a yolk protein precursor, has been implicated in mediating the long lifespan and high fecundity of queen bees. To determine if Vg-like proteins can regulate lifespan in insects generally, we examined the effects of expression of Apis Vg and Drosophila CG31150 (a Vg-like gene recently identified as cv-d) on Drosophila melanogaster lifespan and fecundity using the RU486-inducible GeneSwitch system. For all genotypes tested, overexpression of Vg and CG31150 decreased Drosophila lifespan and did not affect total or age-specific fecundity. We also detected an apparent effect of the GeneSwitch system itself, wherein RU486 exposure (or the GAL4 expression it induces) led to a significant increase in longevity and decrease in fecundity in our fly strains. This result is consistent with the pattern reported in a recent meta-analysis of Drosophila aging studies, where transgenic constructs of the UAS/GAL4 expression system that should have no effect (e.g. an uninduced GeneSwitch) significantly extended lifespan in some genetic backgrounds. Our results suggest that Vg-family genes are not major regulators of Drosophila life history traits, and highlight the importance of using appropriate controls in aging studies.

scholarly journals Conversion of lacZ Enhancer Trap Lines to GAL4 Lines Using Targeted Transposition in Drosophila melanogaster

Genetics ◽  
1999 ◽  
Vol 151 (3) ◽  
pp. 1093-1101 ◽  
Author(s):  
Katharine J Sepp ◽  
Vanessa J Auld
Keyword(s):  
Expression System ◽  
Enhancer Trap ◽  
P Element ◽  
Specific Expression ◽  
Complete Replacement ◽  
New Genes ◽  
Biological Studies ◽  
Gal4 Expression ◽  
Element Mobilization ◽  
Low Efficiency

Abstract Since the development of the enhancer trap technique, many large libraries of nuclear localized lacZ P-element stocks have been generated. These lines can lend themselves to the molecular and biological characterization of new genes. However they are not as useful for the study of development of cellular morphologies. With the advent of the GAL4 expression system, enhancer traps have a far greater potential for utility in biological studies. Yet generation of GAL4 lines by standard random mobilization has been reported to have a low efficiency. To avoid this problem we have employed targeted transposition to generate glial-specific GAL4 lines for the study of glial cellular development. Targeted transposition is the precise exchange of one P element for another. We report the successful and complete replacement of two glial enhancer trap P[lacZ, ry+] elements with the P[GAL4, w+] element. The frequencies of transposition to the target loci were 1.3% and 0.4%. We have thus found it more efficient to generate GAL4 lines from preexisting P-element lines than to obtain tissue-specific expression of GAL4 by random P-element mobilization. It is likely that similar screens can be performed to convert many other P-element lines to the GAL4 system.

scholarly journals Development and testing of a novel Killer-Rescue self-limiting gene drive system in Drosophila melanogaster

2019 ◽  
Author(s):  
Sophia H. Webster ◽  
Michael R. Vella ◽  
Maxwell J. Scott
Keyword(s):  
Drosophila Melanogaster ◽  
Core Promoter ◽  
Expression System ◽  
Drive System ◽  
Gene Drive ◽  
Drosophila Suzukii ◽  
Population Replacement ◽  
Gal4 Expression ◽  
Rescue System ◽  
Release Ratio

AbstractWe report the development and laboratory testing of a novel Killer-Rescue (K-R) self-limiting gene drive system in Drosophila melanogaster. This K-R system utilizes the well-characterized Gal4/UAS binary expression system and the Gal4 inhibitor, Gal80. Three killer (K) lines were tested; these used either an autoregulated UAS-Gal4 or UAS-Gal4 plus UAS-hid transgene. One universal rescue (R) line was used, UAS-Gal80, to inhibit Gal4 expression. The K lines are lethal and cause death in the absence of R. We show that Gal4 RNA levels are high in the absence of R. Death is possibly due to transcriptional squelching from high levels of Gal4. When R is present, Gal4 activation of Gal80 would lead to inhibition of Gal4 and prevent overexpression. With a single release ratio of 2:1 engineered K-R to wildtype, we find that K drives R through the population while the percent of wild type individuals decreases each generation. The choice of core promoter for a UAS-Gal4 construct strongly influences the K-R system. With the strong hsp70 core promoter, K was very effective but was quickly lost from the population. With the weaker DSCP core promoter, K persisted for longer allowing the frequency of individuals with at least one copy of R to increase to over 98%. This simple gene drive system could be readily adapted to other species such as mosquito disease vectors for driving anti-viral or anti-parasite genes.SignificanceHere we report the development and testing of a novel self-limiting gene drive system, Killer-Rescue, in Drosophila melanogaster. This system is composed of an auto-regulated Gal4 Killer (K) and a Gal4-activated Gal80 Rescue (R). Overexpression of Gal4 is lethal but in the presence of R, activation of Gal80 leads to much lower levels of Gal4 and rescue of lethality. We demonstrate that with a single 2:1 engineered to wildtype release, more than 98% of the population carry R after eight generations. We discuss how this Killer-Rescue system may be used for population replacement in a human health pest, Aedes aegypti, or for population suppression in an agricultural pest, Drosophila suzukii.

scholarly journals New fluorescent protein reporters for use with thedrosophila gal4 expression system and for vital detection of balancer chromosomes

genesis ◽  
2002 ◽  
Vol 34 (1-2) ◽  
pp. 135-138 ◽  
Author(s):  
Marc S. Halfon ◽  
Stephen Gisselbrecht ◽  
Jun Lu ◽  
Beatriz Estrada ◽  
Haig Keshishian ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Expression System ◽  
Gal4 Expression ◽  
Balancer Chromosomes

scholarly journals AGES: An auxin-inducible, GAL4-compatible, gene expression system for Drosophila

2021 ◽  
Author(s):  
Colin D. McClure ◽  
Amira Hassan ◽  
Aneisha Duggal ◽  
Chee Ying Sia ◽  
Tony D. Southall
Keyword(s):  
Gene Expression ◽  
Expression System ◽  
Cost Effective ◽  
Water Soluble ◽  
Model Organisms ◽  
Temperature Sensitive ◽  
Gal4 Driver ◽  
Gene Expression System ◽  
Gal4 Expression ◽  
Inducible Systems

AbstractThe ability to control transgene expression, both spatially and temporally, is essential for studying model organisms. In Drosophila, spatial control is primarily provided by the GAL4/UAS system, whilst temporal control relies on a temperature-sensitive GAL80 (which inhibits GAL4) and drug-inducible systems. However, these are not ideal. Shifting temperature can impact on many physiological and behavioural traits, and the current drug-inducible systems are either leaky, toxic, incompatible with existing GAL4-driver lines, or do not generate effective levels of expression. Here we describe the Auxin-inducible Gene Expression System (AGES). AGES relies on the auxin-dependent degradation of a ubiquitously expressed GAL80, and therefore, is compatible with existing GAL4-driver lines. Water-soluble auxin is added to fly food at a low, non-lethal, concentration, which induces expression comparable to uninhibited GAL4 expression. The system works in both larvae and adults, providing a stringent, non-lethal, cost-effective, and convenient method for temporally controlling GAL4 activity in Drosophila.

High Level Expression of Active Human Prothrombin in a Vaccina Virus Expression System

1992 ◽  
Vol 68 (02) ◽  
pp. 119-124 ◽  
Author(s):  
F G Falkner ◽  
P L Turecek ◽  
R T A MacGillivray ◽  
W Bodemer ◽  
F Scheiflinger ◽  
...  
Keyword(s):  
Vaccinia Virus ◽  
Cell Lines ◽  
Expression System ◽  
Human Cell Line ◽  
Cell System ◽  
Time Saving ◽  
Expression Levels ◽  
Mammalian Cell Lines ◽  
Cell Line Hela ◽  
Virus Expression

SummaryWe have worked out an efficient and time saving procedure for the expression of recombinant human prothrombin. The glycoprotein was expressed in the vaccinia virus expression system in several mammalian cell lines. The kidney cell lines Vero and BHK and the human cell line Hela were found to efficiently secrete prothrombin. Expression levels of 3–4 µg of factor II per 106 cells per day corresponding to 18–23 mU per 106 cells per day were achieved. Since the expression levels obtained with the vaccinia virus/Vero cell system were comparable to those obtained in amplified transformed CHO cells it provides an alternative system for the efficient expression of human prothrombin and may allow to further elucidate structure-function relationships of (pro)thrombin and its various effectors.

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