scholarly journals P-lacW insertional mutagenesis on the second chromosome of Drosophila melanogaster: isolation of lethals with different overgrowth phenotypes.

Genetics ◽  
1993 ◽  
Vol 135 (1) ◽  
pp. 71-80 ◽  
Author(s):  
T Török ◽  
G Tick ◽  
M Alvarado ◽  
I Kiss
Keyword(s):  
Drosophila Melanogaster ◽  
In Situ Hybridization ◽  
Insertional Mutagenesis ◽  
Imaginal Discs ◽  
Larval Period ◽  
P Element ◽  
Lymph Glands ◽  
Hematopoietic Organs

Abstract A single P-element insertional mutagenesis experiment was carried out for the second chromosome of Drosophila melanogaster using the P-lacW transposon. Out of 15,475 insertions on the second chromosome, 2,308 lethal and 403 semilethal mutants (altogether 2,711) were recovered. After eliminating clusters, 72% of the mutants represent independent insertions. Some of the mutants with larval, prepupal or pupal lethal phases have a prolonged larval period and show gradual overgrowth of the imaginal discs, brain and/or the hematopoietic organs (lymph glands). In this paper, 16 overgrowth mutants are described. As revealed by in situ hybridization, none of the mutations corresponds to any of the previously known overgrowth mutations on the second chromosome.

Transformation of Drosophila melanogaster with a suppressor tRNA gene from Schizosaccharomyces pombe

Genome ◽  
1988 ◽  
Vol 30 (2) ◽  
pp. 211-217 ◽  
Author(s):  
Charles M. Molnar ◽  
Tove Reece ◽  
James A. Williams ◽  
John B. Bell
Keyword(s):  
Drosophila Melanogaster ◽  
Salivary Gland ◽  
In Situ Hybridization ◽  
Schizosaccharomyces Pombe ◽  
Southern Hybridization ◽  
Trna Gene ◽  
P Element ◽  
Suppressor Trna ◽  
Good Agreement

P-element mediated transformation was utilized to introduce a suppressor tRNA gene [Formula: see text] from Schizosaccharomyces pombe into Drosophila melanogaster. Thirteen independently transformed lines were characterized as to the number and cytological locations of the transposons. It was ascertained that the suppressor tRNA gene of interest was introduced into each transformed strain. The helper P element used (pπ25.1) allows further transposition to occur, and it was determined that from one to seven copies of the heterologous [Formula: see text] gene per strain were present among the respective transformed strains. The number of transposons per transformed line was established by in situ hybridization to salivary gland chromosomes as well as by Southern hybridization analyses and there was good agreement in the totals determined by these two techniques.Key words: Drosophila, Schizosaccharomyces, tRNA suppressor, transformation, transposon.

Transcription of the segment-polarity gene wingless in the imaginal discs of Drosophila, and the phenotype of a pupal-lethal wg mutation

Development ◽  
1988 ◽  
Vol 102 (3) ◽  
pp. 489-497 ◽  
Author(s):  
N.E. Baker
Keyword(s):  
In Situ Hybridization ◽  
Imaginal Discs ◽  
Apical Cytoplasm ◽  
Segment Polarity ◽  
Disc Cells ◽  
Segment Polarity Gene ◽  
Drosophila Embryos ◽  
Polarity Gene

Wingless (wg) is a segment-polarity gene in Drosophila which is related to the murine proto-oncogene int1. In Drosophila embryos, wg transcription defines part of each parasegment. In situ hybridization shows that wg is also expressed in the imaginal discs which give rise to the adult during metamorphosis. Transcripts are localized in the apical cytoplasm of disc cells, and accumulate in different patterns in dorsal and ventral discs. The wgCX3 mutation produces morphological defect in the adult structures derived from these imaginal discs. The results show that wg is involved in the development of the adult, as well as the embryo, but that the imaginal discs do not express this segment-polarity gene in an identical pattern to the embryonic segments.

Quantitative in situ hybridization of ribosomal RNA species to polytene chromosomes of Drosophila melanogaster

1977 ◽  
Vol 115 (3) ◽  
pp. 539-563 ◽  
Author(s):  
Paul Szabo ◽  
Robert Elder ◽  
Dale M. Steffensen ◽  
Olke C. Uhlenbeck
Keyword(s):  
Drosophila Melanogaster ◽  
In Situ Hybridization ◽  
Ribosomal Rna ◽  
Polytene Chromosomes

scholarly journals Chromosome interactions in P-M dysgenic male recombination of Drosophila melanogaster

1992 ◽  
Vol 60 (3) ◽  
pp. 165-174
Author(s):  
P. Eggleston ◽  
K. A. Exley
Keyword(s):  
Drosophila Melanogaster ◽  
Recombination Breakpoint ◽  
P Element ◽  
Chromosome 2 ◽  
Male Recombination ◽  
P Elements ◽  
Element Array ◽  
Recombination Breakpoints ◽  
Map Distance

SummaryThe frequency, distribution and structure of P elements on the second and third chromosomes of Texas 1, a wild-type inbred strain of Drosophila melanogaster, were investigated by in situ hybridization. These autosomes were isolated individually and used as P-element donors to study the frequency and distribution of male recombination events generated on recipient chromosomes which were originally devoid of P sequences. The P-element array of chromosome 2 was shown to generate higher male recombination frequencies on chromosome 3 than vice versa, despite having fewer P factors and fewer P elements in general. This is likely to be due to the presence and distribution of specific P-deletion derivatives, which vary in their ability to repress P mobility. The male recombination generated on recipient chromosomes is associated with the insertion of donated P sequences, but only in a small minority of cases could a novel P-element site be detected at, or near, the recombination breakpoint. The majority of such breakpoints appear to be associated either with unsuccessful P insertion, or with the action of P transposase attracted by P elements newly inserted elsewhere on the recipient chromosome. Recent evidence also suggests that a small proportion of the breakpoints may be associated with the action of P transposase alone. Male recombination breakpoints appear to be distributed effectively at random along the recipient autosomes, and their frequency of occurrence was shown to correlate with the physical length of DNA available between markers, as revealed by the polytene map distance.

HCR RNA-FISH protocol for the whole-mount brains of Drosophila and other insects v1

2021 ◽  
Author(s):  
Amanda A. G. Ferreira ◽  
Bogdan Sieriebriennikov ◽  
Hunter Whitbeck
Keyword(s):  
Drosophila Melanogaster ◽  
In Situ Hybridization ◽  
Fluorescent In Situ Hybridization ◽  
Hybridization Chain Reaction ◽  
Chain Reaction ◽  
Alexa Fluor ◽  
Rna Fish ◽  
Whole Mount

This is a protocol to perform RNA fluorescent in situ hybridization (RNA-FISH) using hybridization chain reaction (HCR) on whole-mount samples of the brains of the fly Drosophila melanogaster and other insects, e.g. the jumping ant Harpegnathos saltator. Probes and HCR reagents are purchased from Molecular Instruments. This protocol is loosely based on the "generic sample in solution" protocol published by Molecular Instruments. Our modifications include the description of fixation conditions, counterstaining by Hoechst, and altered washes. Additionally, we use larger concentrations of probes and hairpins following the protocol described by Younger, Herre et al. 2020. We have successfully employed this protocol to stain insect brains with up to 4 different probe sets simultaneously (hairpins conjugated with Alexa Fluor 488, 546, 496, and 647).

Drosophila melanogaster homologs of the raf oncogene

1987 ◽  
Vol 7 (6) ◽  
pp. 2134-2140
Author(s):  
G E Mark ◽  
R J MacIntyre ◽  
M E Digan ◽  
L Ambrosio ◽  
N Perrimon
Keyword(s):  
Drosophila Melanogaster ◽  
In Situ Hybridization ◽  
Steady State ◽  
Kinase Domain ◽  
Early Embryogenesis ◽  
The Other ◽  
Chromosome 2 ◽  
Related Gene ◽  
Steady State Levels

A murine v-raf probe, representing the kinase domain, was used to identify two unique loci in Drosophila melanogaster DNA. The most closely related to v-raf was mapped by in situ hybridization to position 2F5-6 (Draf-1) on the X chromosome, whereas the other raf-related gene (Draf-2) was found at position 43A2-5 on chromosome 2. The nucleotide and amino acid homologies of Draf-1 to the kinase domain of v-raf are 61 and 65%, respectively. The large amount of a 3.2-kilobase Draf-1 transcript detected in eggs as a maternal message decreases during embryonic development, and significant steady-state levels are observed throughout the remainder of morphogenesis. We speculate that the Draf-1 locus plays an important role in early embryogenesis.

scholarly journals The fitness consequences of P element insertion in Drosophila melanogaster

1988 ◽  
Vol 52 (1) ◽  
pp. 17-26 ◽  
Author(s):  
Walter F. Eanes ◽  
Cedric Wesley ◽  
Jody Hey ◽  
David Houle ◽  
James W. Ajioka
Keyword(s):  
Drosophila Melanogaster ◽  
Relative Number ◽  
African Population ◽  
P Element ◽  
Insertion Event ◽  
Element Insertion ◽  
Lethal Mutations ◽  
Fitness Consequences ◽  
Per Se

SummaryIn this study we estimate the frequency at which P-element insertion events, as identified by in situ hybridization, generate lethal and mild viability mutations. The frequency of lethal mutations generated per insertion event was 0·004. Viability dropped an average of 1% per insertion event. Our results indicate that it is deletions and rearrangements resulting from the mobilization of P elements already in place and not the insertions per se that cause the drastic effects on viability and fitness observed in most studies of P–M dysgenesis-derived mutations. Elements of five other families (I, copia, 412, B104, and gypsy) were not mobilized in these crosses. Finally, we contrast the density of P elements on the X chromosome with the density on the four autosomal arms in a collection of thirty genomes from an African population. The relative number of P elements on the X chromosome is too high to be explained by either a hemizygous selection or a neutrality model. The possible reasons for the failure to detect selection are discussed.

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