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A Spradling

Publications and source records attributed to A Spradling.

15 recordsLinked to original sources
PubMed1988

Insertional mutagenesis of the Drosophila genome with single P elements.

A versatile genetic method for identifying and cloning Drosophila melanogaster genes affecting any recognizable phenotype is described. Strains are constructed in which the insertion of a single P transposable element has caused a new mutation, greatly simplifying the genetic and molecular analysis of the affected gene. Mutagenesis is initiated by crossing two strains, each of which contains a specially designed P element. One element (jumpstarter), encoding P element transposase, efficiently mobilizes the second nonautonomous transposon (mutator), whose structure facilitates selection and cloning of new insertion mutations. Random mutator transpositions are captured in individual stocks that no longer contain jumpstarter, where they remain stable. This method was used to construct 1300 single P element insertion stocks which were then screened for recessive mutations. A library of single-element insertion strains will allow the structure and function of Drosophila genes to be readily correlated, and should have many other applications in Drosophila molecular genetics.

Animals↗
PubMed1986

Replication and expression of an X-linked cluster of Drosophila chorion genes.

Two 80- to 100-kb chromosomal replicons containing clustered chorion genes amplify in the ovarian follicle cells during the final 22 hr of Drosophila oogenesis. We have studied the relationship between amplification and transcription within one of these domains, located at 7E10-7F3,4 on the X chromosome. A tandem cluster of six genes, encoding chorion structural proteins s36-1, s38-1, and four putative minor chorion protein mRNAs, was mapped in the central 18 kb of the amplified domain, a region showing the highest levels of amplification. The regions both proximal and distal to this gene cluster, where lower levels of amplification occur, were also transcribed in ovary, but mRNAs produced specifically during choriogenesis were not detected. Thus, differences in amplification do not appear to modulate differential RNA accumulation. Instead, the gradient of amplification observed in egg chamber DNA may simply reflect the mechanism of amplification. In the female sterile mutation, In(1)ocelliless, a chromosomal rearrangement separates the central gene cluster into two parts, only one of which retains the capacity to amplify. Genes located within the unamplified portion of the ocelliless chromosome were expressed at the appropriate time during oogenesis, but at a 5- to 10-fold reduced level of RNA per gene. Thus neither cluster integrity nor amplification are required for the normal developmental program of gene expression within the cluster.

Animals↗
PubMed1985

DNA sequence of a 3.8 kilobase pair region controlling Drosophila chorion gene amplification.

During Drosophila oogenesis, two clusters of chorion genes and their flanking DNA sequences undergo amplification in the ovarian follicle cells. Amplification results from repeated rounds of initiation and bidirectional replication within the chorion gene regions, possibly from a single origin, producing nested replication forks. Previously we have shown that following reintroduction into the Drosophila genome, a specific 3.8 kilobase pair DNA segment from the amplified third chromosome domain could induce developmentally regulated amplification at its site of insertion. Here we present the complete nucleotide sequence of this "amplification control element" and of genes encoding the chorion structural proteins s18-1 and s15-1, which are contained within it. Sequences that may be involved in the regulation of chorion gene amplification and expression are identified.

Animals↗
PubMed1975

Two very different components of messenger RNA in an insect cell line.

The messenger RNA lifetimes have been measured in a cell line derived from an invertebrate source, the mosquito Aedes albopictus. The experiments were made possible by a new technique for obtaining undegraded cytoplasmic RNA from cells with high endogenous nuclease levels. There are two components to the decay kinetics of Aedes mRNA. The major fraction of the steady state message population has a half-life of 20 hr which is, as in mammalian cells, comparable to the cell generation time. The short-lived component turns over very rapidly with a half-life estimated to be about 1.2 hr. The difference in lifetime between the short and long-lived components is about 15 fold in these cells, compared to 3-4 fold in mammalian cells. This may reflect the need for a more responsive mRNA regulating system in poikilothermic organisms. The great disparity between the principle messenger lifetimes permits a more definite assignment of a two component behavior to message decay. The data in the case of mammalian cells could not rule out a family of intermediate lifetimes. The long-lived mRNA has a much smaller average sedimentation value than the short-lived material. The effect is similar to, but much larger than, that seen in mammalian cells. Although the lifetime difference is much greater in the insect cells than in human (HeLa) cells, the fast and slow components comprise about the same proportion of the steady state mRNA population: 30 percent and 70 percent, respectively.

Aedes↗
PubMed1975

Analysis of drosophila mRNA by in situ hybridization: sequences transcribed in normal and heat shocked cultured cells.

Messenger RNA transcribed in cultured Drosophila cells adapted for growth under conditions permitting labeling to high specific acitivty has been analyzed by the technique of in situ hybridization. Poly(A)-containing cytoplasmic RNA binds specifically and reproducibly to about 50 bands in the salivary gland polytene chromosomes. In addition heavy labeling of the beta-heterochromatin associated with each of the chromosome arms is observed. The species which are detected probably belong to the more abundant classes of RNA. When the cultured Drosophila cells are subjected to heat shock immediately before labeling with 3H-uridine, there is a drastic alteration in the pattern of gene transcription detected by in situ hybridization. Most of the mRNA synthesis which could be detected in the normal cell is shut off. Newly synthesized RNA hybridizes strongly to seven new sites which do not bind mRNA from control cells. The new loci correspond almost exactly to the regions of Drosophila polytene chromosomes which puff when intact larvae are subjected to an identical heat treatment.

Animals↗