Molecular characterization of the cinnabar region of Drosophila melanogaster: Identification of the cinnabar transcription unit

Warren, William D.; Palmer, Stephanie O.; Howells, A. J.

doi:10.1007/bf00057589

Cited by 43 publications

(34 citation statements)

References 67 publications

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“…The transformation vector pH [cn], which includes the cn ϩ gene fragment from D. melanogaster (13) flanked by the terminal inverted repeats of the Hermes transposon, was developed as follows. All Hermes fragments have been described by Warren et al (14).…”

Section: Methodsmentioning

confidence: 99%

“…The resulting plasmid contained unique BamHI and SalI sites between the two arms into which the LacZ alpha peptide of pBluescript SK (Stratagene) was inserted as a BglII͞XhoI-digested fragment produced by PCR by using the primers, 5Ј-GCAGATCTAT-CAGGGCGATGGCC-3Ј and 5Ј-GCCTCGAGCTGGCAC-GACAGGTTTCCCG-3Ј. This plasmid, pH [LacZ], was modified further by cutting with XbaI and SacII, and inserting a 4.7-kb XbaI͞SacII fragment of D. melanogaster genomic DNA containing a wild-type copy of the cn gene (13). The resulting plasmid pH[cn] and the transposase encoding helper plasmid pHSHH1.9 (15) were purified through a cesium chloride gradient by ultracentrifugation, washed five times with isopropanol saturated with NaCl, and precipitated in three volumes of 70% ethanol.…”

Section: Methodsmentioning

confidence: 99%

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Stable transformation of the yellow fever mosquito, Aedes aegypti , with the Hermes element from the housefly

Jasinskiene

Coates²,

Benedict³

et al. 1998

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

331

203

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The mosquito Aedes aegypti is the world's most important vector of yellow fever and dengue viruses. Work is currently in progress to control the transmission of these viruses by genetically altering the capacity of wild Ae. aegypti populations to support virus replication. The germline transformation system reported here constitutes a major advance toward the implementation of this control strategy. A modified Hermes transposon carrying a 4.7-kb fragment of genomic DNA that includes a wild-type allele of the Drosophila melanogaster cinnabar (cn) gene was used to transform a white-eyed recipient strain of Ae. aegypti. Microinjection of preblastoderm mosquito embryos with this construct resulted in 50% of the emergent G 0 adults showing some color in their eyes. Three transformed families were recovered, each resulting from an independent insertion event of the cn ؉ -carrying transposon. The cn ؉ gene functioned as a semidominant transgene and segregated in Mendelian ratios. Hermes shows great promise as a vector for efficient, heritable, and stable transformation of this important mosquito vector species.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Stable transformation of the yellow fever mosquito, Aedes aegypti , with the Hermes element from the housefly

Jasinskiene

Coates²,

Benedict³

et al. 1998

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

331

203

View full text Add to dashboard Cite

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“…The primary ommochrome precursor is tryptophan that is converted into 3-hydroxykynurenine by three enzymes encoded by vermilion, kynurenine formamidase and cinnabar (Searles and Voelker, 1986;Searles et al, 1990;Warren et al, 1996). Subsequently, 3-hydroxykynurenine is incorporated into pigment granules by the heterodimeric ABC (ATP-binding cassette) transporters encoded by scarlet and white (Tearle et al, 1989;Pepling and Mount, 1990).…”

Section: Introductionmentioning

confidence: 99%

Positional cloning of a Bombyx pink-eyed white egg locus reveals the major role of cardinal in ommochrome synthesis

Osanai-Futahashi

Futahashi

et al. 2015

Heredity

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Ommochromes are major insect pigments involved in coloration of compound eyes, eggs, epidermis and wings. In the silkworm Bombyx mori, adult compound eyes and eggs contain a mixture of the ommochrome pigments such as ommin and xanthommatin. Here, we identified the gene involved in ommochrome biosynthesis by positional cloning of B. mori egg and eye color mutant pink-eyed white egg (pe). The recessive homozygote of pe has bright red eyes and white or pale pink eggs instead of a normal dark coloration due to the decrease of dark ommochrome pigments. By genetic linkage analysis, we narrowed down the pe-linked region to~258 kb, containing 17 predicted genes. RNA sequencing analyses showed that the expression of one candidate gene, the ortholog of Drosophila haem peroxidase cardinal, coincided with egg pigmentation timing, similar to other ommochrome-related genes such as Bm-scarlet and Bm-re. In two pe strains, a common missense mutation was found within a conserved motif of B. mori cardinal homolog (Bm-cardinal). RNA interference-mediated knockdown and transcription activatorlike effector nuclease (TALEN)-mediated knockout of the Bm-cardinal gene produced the same phenotype as pe in terms of egg, adult eye and larval epidermis coloration. A complementation test of the pe mutant with the TALEN-mediated Bm-cardinaldeficient strain showed that the mutant phenotype could not be rescued, indicating that Bm-cardinal is responsible for pe. Moreover, knockdown of the cardinal homolog in Tribolium castaneum also induced red compound eyes. Our results indicate that cardinal plays a major role in ommochrome synthesis of holometabolous insects.

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“…The transposable elements P, Minos, piggyBac, and hermes have, among others, all been successfully used to drive integration of exogenous DNA into the fruit fly genome (8 -11). At the same time, the use of an array of visible selectable markers, such as white, rosy, cinnabar, and more recently EGFP (12)(13)(14)(15), has greatly facilitated the screening procedure and allowed complex functional studies of genes and their promoter regions. Germline transformation of A. stephensi mosquitoes is anticipated to open a wide range of applications to explore the genome of these important disease vectors when sustained by a parallel development of appropriate molecular tools.…”

mentioning

confidence: 99%

piggyBac-mediated Germline Transformation of the Malaria Mosquito Anopheles stephensi Using the Red Fluorescent Protein dsRED as a Selectable Marker

Nolan¹,

Bower²,

Brown

et al. 2002

Journal of Biological Chemistry

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It is estimated that every year malaria infects ϳ300 million people and accounts for the death of 2 million individuals. The Plasmodium parasites that cause malaria in humans are transmitted exclusively by mosquito species belonging to the Anopheles genus. The recent development of a gene transfer technology for Anopheles stephensi mosquitoes, using the Minos transposable element marked with the enhanced green fluorescent protein EGFP (Catteruccia, F., Nolan, T., Loukeris, T. G., Blass, C., Savakis, C., Kafatos, F. C., and Crisanti, A. (2000) Nature 405, 959 -962), provides now a powerful tool to investigate the role of mosquito molecules involved in the interaction with the malaria parasite. Such technology, when further developed with additional markers and transposable elements, will be invaluable for analyzing the biology of the vector and for developing malaria-resistant mosquitoes to be used as a tool to control malaria transmission in the field. We report here the germline transformation of A. stephensi mosquitoes using a piggyBac-based transposon to drive integration of the gene encoding for the red fluorescent protein dsRED. A. stephensi embryos were injected with transformation vector pPBRED containing the dsRED marker cloned within the arms of piggyBac. Microscopic analysis of G 1 larvae revealed the presence of seven fluorescent phenotypes whose different molecular origins were confirmed by Southern blotting analysis. Sequencing of the insertion sites in two lines demonstrated that integrations had occurred at TTAA nucleotides in accordance with piggyBac-mediated transpositions.The recent development of an efficient gene transfer technology for Anopheles stephensi mosquitoes, achieved by using a Minos-based transposon (4) loaded with the EGFP 1 selectable marker (1), has expanded the possibility of studying the genetics of human malaria vectors at the functional level. Although EGFP has proven to be an invaluable visible marker for identifying transformed individuals in different insect species (1, 5-7), the availability of only this selectable marker limits the range of applications of a gene transfer technology in malaria vectors. New molecular tools are now needed to exploit fully the potential of germline transformation for Anopheles mosquitoes, with the aim of unraveling the interactions between host molecules and Plasmodium parasites, as well as performing functional studies such as transposon tagging and enhancer trapping. Ultimately this technology could be used to develop transgenic mosquitoes with a nonpermissive phenotype for parasite development. These mosquitoes could be used in malaria control programs with the aim to replace the permissive wild type vectors.As shown in Drosophila melanogaster, the availability of various molecular and genetic tools to achieve germline transformation has contributed tremendously to our understanding of the fruit fly biology, leading to the identification of hundreds of genes involved in development, immunity, tissue modeling, and embryogenesis. The tran...

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Molecular characterization of the cinnabar region of Drosophila melanogaster: Identification of the cinnabar transcription unit

Cited by 43 publications

References 67 publications

Stable transformation of the yellow fever mosquito, Aedes aegypti , with the Hermes element from the housefly

Stable transformation of the yellow fever mosquito, Aedes aegypti , with the Hermes element from the housefly

Positional cloning of a Bombyx pink-eyed white egg locus reveals the major role of cardinal in ommochrome synthesis

piggyBac-mediated Germline Transformation of the Malaria Mosquito Anopheles stephensi Using the Red Fluorescent Protein dsRED as a Selectable Marker

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