Isolation of mutations in the Drosophila homologues of the human Neurofibromatosis 2 and yeast CDC42 genes using a simple and efficient reverse-genetic method

Reverse genetic analysis in Drosophila has been greatly aided by a growing collection of lethal P transposable element insertions that provide molecular tags for the identification of essential genetic loci. However, because the screens performed to date primarily have generated autosomal P-element...

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Bibliographic Details
Published in:Genetics (Austin) 1997-05, Vol.146 (1), p.245-252
Main Authors: Fehon, R.G. (Duke University, Durham, NC.), Oren, T, LaJeunesse, D.R, Melby, T.E, McCartney, B.M
Format: Article
Language:English
Subjects:
ADN
Animals
cdc42 GTP-Binding Protein, Saccharomyces cerevisiae
Cell Cycle Proteins - genetics
CHROMOSOME
CHROMOSOMES
COMPLEMENTARY DNA
COMPLEMENTATION
COMPLEMENTATION GROUPS
COSMID TRANSGENES
Cosmids
CROMOSOMAS
DESARROLLO EMBRIONARIO
DEVELOPPEMENT EMBRYONNAIRE
DNA
Drosophila
Drosophila - genetics
DROSOPHILA MELANOGASTER
EMBRYONIC DEVELOPMENT
ENSAYO
Female
GENE
GENE LETAL
GENES
GENES LETALES
Genes, Neurofibromatosis 2
GENETIC ANALYSIS
Genetic Complementation Test
GENETICA
GENETICS
GENETIQUE
GTP-Binding Proteins - genetics
HEMIZYGOSITY
Humans
Insects
Investigations
LETHAL GENES
Male
Membrane Proteins - genetics
MERLIN GENE
Molecular Sequence Data
MUTACION
MUTAGENESIS
Mutagenesis, Insertional
MUTATION
Neurofibromin 2
NUCLEIC PROBES
POLYTENE CHROMOSOMES
REVERSE GENETIC ANALYSIS
SCREENING
SONDAS NUCLEICAS
SONDE NUCLEIQUE
TESTAGE
TESTING
Tumors
Yeast
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Description
Summary:Reverse genetic analysis in Drosophila has been greatly aided by a growing collection of lethal P transposable element insertions that provide molecular tags for the identification of essential genetic loci. However, because the screens performed to date primarily have generated autosomal P-element insertions, this collection has not been as useful for performing reverse genetic analysis of X-linked genes. We have designed a reverse genetic screen that takes advantage of the hemizygosity of the X chromosome in males together with a cosmid-based transgene that serves as an autosomally linked duplication of a small region of the X chromosome. The efficacy and efficiency of this method is demonstrated by the isolation of mutations in Drosophila homologues of two well-studied genes, the human Neurofibromatosis 2 tumor suppressor and the yeast CDC42 gene. The method we describe should be of general utility for the isolation of mutations in other X-linked genes, and should also provide an efficient method for the isolation of new allcles of existing X-linked or autosomal mutations in Drosophila.
ISSN:0016-6731
1943-2631
1943-2631
DOI:10.1093/genetics/146.1.245