Fluorescence in situ Hybridization with Bacterial Artificial Chromosomes (BACs) to Mitotic Heterochromatin of Drosophila

Accardo, Maria Carmela; Dimitri, Patrizio

doi:10.1007/978-1-60761-789-1_30

Cited by 3 publications

(2 citation statements)

References 11 publications

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“…BAC DNA was prepared by the standard alkaline lysis method. To limit bias in the interpretation of FISH results, BAC preparations were assigned a code number, and identical coded samples were delivered to the Dimitri laboratory for mitotic FISH (Brizuela et al 1994) was performed as described in Accardo and Dimitri (2010).…”

Section: Cytogenetic Mapping Of Bacsmentioning

confidence: 99%

The Release 6 reference sequence of the Drosophila melanogaster genome

et al. 2015

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Drosophila melanogaster plays an important role in molecular, genetic, and genomic studies of heredity, development, metabolism, behavior, and human disease. The initial reference genome sequence reported more than a decade ago had a profound impact on progress in Drosophila research, and improving the accuracy and completeness of this sequence continues to be important to further progress. We previously described improvement of the 117-Mb sequence in the euchromatic portion of the genome and 21 Mb in the heterochromatic portion, using a whole-genome shotgun assembly, BAC physical mapping, and clone-based finishing. Here, we report an improved reference sequence of the single-copy and middle-repetitive regions of the genome, produced using cytogenetic mapping to mitotic and polytene chromosomes, clone-based finishing and BAC fingerprint verification, ordering of scaffolds by alignment to cDNA sequences, incorporation of other map and sequence data, and validation by whole-genome optical restriction mapping. These data substantially improve the accuracy and completeness of the reference sequence and the order and orientation of sequence scaffolds into chromosome arm assemblies. Representation of the Y chromosome and other heterochromatic regions is particularly improved. The new 143.9-Mb reference sequence, designated Release 6, effectively exhausts clone-based technologies for mapping and sequencing. Highly repeat-rich regions, including large satellite blocks and functional elements such as the ribosomal RNA genes and the centromeres, are largely inaccessible to current sequencing and assembly methods and remain poorly represented. Further significant improvements will require sequencing technologies that do not depend on molecular cloning and that produce very long reads.

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Section: Cytogenetic Mapping Of Bacsmentioning

confidence: 99%

The Release 6 reference sequence of the Drosophila melanogaster genome

et al. 2015

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“…Mitotic chromosome preparations were stained with DAPI. FISH procedures were performed as reported (62). Biotinylated anti-avidin antibodies were used to amplify the hybridization signal.…”

Section: Rna Preparation and Rt-pcr Experimentsmentioning

confidence: 99%

Functional evidence that a recently evolved Drosophila sperm-specific gene boosts sperm competition

Yeh

Chan

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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In many species, both morphological and molecular traits related to sex and reproduction evolve faster in males than in females. Ultimately, rapid male evolution relies on the acquisition of genetic variation associated with differential reproductive success. Many newly evolved genes are associated with novel functions that might enhance male fitness. However, functional evidence of the adaptive role of recently originated genes in males is still lacking. The Sperm dynein intermediate chain multigene family, which encodes a Sperm dynein intermediate chain presumably involved in sperm motility, originated from complex genetic rearrangements in the lineage that leads to Drosophila melanogaster within the last 5.4 million years since its split from Drosophila simulans. We deleted all the members of this multigene family resident on the X chromosome of D. melanogaster by chromosome engineering and found that, although the deletion does not result in a reduction of progeny number, it impairs the competence of the sperm in the presence of sperm from wildtype males. Therefore, the Sperm dynein intermediate chain multigene family contributes to the differential reproductive success among males and illustrates precisely how quickly a new gene function can be incorporated into the genetic network of a species.Faster-male evolution | male fertility

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Cytological heterogeneity of heterochromatin among 10 sequenced Drosophila species

et al. 2022

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In Drosophila chromosomal rearrangements can be maintained and are associated with karyotypic variability among populations from different geographic localities. The abundance of variability in gene arrangements among chromosomal arms is even greater when comparing more distantly related species and the study of these chromosomal changes has provided insights into the evolutionary history of species in the genus. Additionally, the sequencing of genomes of several Drosophila species has offered the opportunity to establish the global pattern of genomic evolution, at both genetic and chromosomal level. The combined approaches of comparative analysis of syntenic blocks and direct physical maps on polytene chromosomes have elucidated changes in the orientation of genomic sequences and the difference between heterochromatic and euchromatic regions. Unfortunately, the centromeric heterochromatic regions cannot be studied using the cytological maps of polytene chromosomes because they are underreplicated and therefore reside in the chromocenter. In D. melanogaster, a cytological map of the heterochromatin has been elaborated using mitotic chromosomes from larval neuroblasts. In the current work we have expanded on that mapping by producing cytological maps of the mitotic heterochromatin in an additional 10 sequenced Drosophila species. These maps highlight two apparently different paths, for the evolution of the pericentric heterochromatin between the subgenera Sophophora and Drosophila. One path leads towards a progressive complexity of the pericentric heterochromatin (Sophofora) and the other towards a progressive simplification (Drosophila). These maps are also useful for a better understanding how karyotypes have been altered by chromosome arm reshuffling during evolution.

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Fluorescence in situ Hybridization with Bacterial Artificial Chromosomes (BACs) to Mitotic Heterochromatin of Drosophila

Cited by 3 publications

References 11 publications

The Release 6 reference sequence of the Drosophila melanogaster genome

The Release 6 reference sequence of the Drosophila melanogaster genome

Functional evidence that a recently evolved Drosophila sperm-specific gene boosts sperm competition

Cytological heterogeneity of heterochromatin among 10 sequenced Drosophila species

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