We have used a Tol2-derived trapping vector, carrying a hybrid Gal4 gene and a UAS:eGFP reporter cassette, to identify 16 transgenic zebrafish lines expressing the fluorescent marker eGFP in tissue-restricted patterns during development. Most lines show co-expression of eGFP and a hybrid Gal4 transcription activator containing a truncated VP16 domain that facilitate induction of other UAS-transgenes (UAS:RFP). Notably, many of the transgenic lines are expressed in particular areas of the central nervous system, such as the retina. We mapped the genomic positions of most of the activated insertions, and for three retina-specific lines we also demonstrate that eGFP reports the expression of particular endogenous genes. One of the identified zebrafish genes shows expression in ventral retina, and encodes a protein containing a repulsive guidance molecule (RGM) domain, suggesting a role in axonal guidance during optic nerve formation. Among the lines labeling other tissues, three show early co-expression of eGFP and Gal4-VP16 in blood vessels, erythrocytes and other hematopoietic cells. Interestingly, the activated insertion in the erythrocyte line was mapped to a site near the globin cluster on chromosome 3. All the reported lines co-expressing eGFP and the hybrid Gal4 activator may have potential as genetic tools to study developmental processes.
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http://dx.doi.org/10.1016/j.gep.2011.09.001 | DOI Listing |
J Genet Eng Biotechnol
December 2024
Center for Genomics, Helmy Institute for Medical Sciences, Zewail City of Science and Technology, Giza, Egypt; University of Science and Technology, Zewail City of Science and Technology, Giza, Egypt. Electronic address:
The utilization of human cDNA libraries in yeast genetic screens is an approach that has been used to identify novel gene functions and/or genetic and physical interaction partners through forward genetics using yeast two-hybrid (Y2H) and classical cDNA library screens. Here, we summarize several challenges that have been observed during the implementation of human cDNA library screens in Saccharomyces cerevisiae (budding yeast). Upon the utilization of DNA repair deficient-yeast strains to identify novel genes that rescue the toxic effect of DNA-damage inducing drugs, we have observed a wide range of transcripts that could rescue the strains.
View Article and Find Full Text PDFBiomolecules
October 2024
Department of Biomedical Sciences, School of Biological and Environmental Sciences, Kwansei Gakuin University, 1 Gakuen Uegahara, Sanda 669-1330, Hyogo, Japan.
The central complex (CX) plays a key role in many higher-order functions of the insect brain including navigation and activity regulation. Genetic tools for manipulating individual cell types, and knowledge of what neurotransmitters and neuromodulators they express, will be required to gain mechanistic understanding of how these functions are implemented. We generated and characterized split-GAL4 driver lines that express in individual or small subsets of about half of CX cell types.
View Article and Find Full Text PDFJ Biol Chem
November 2024
Waseda Research Institute for Science and Engineering, Waseda University, Shinjuku, Tokyo, Japan; Department of Chemistry and Biochemistry, School of Advanced Science and Engineering, Waseda University, Shinjuku, Tokyo, Japan. Electronic address:
J Biol Chem
August 2024
Laboratorio de Glicómica Funcional y Molecular, Programa de Glicoinmunología, Instituto de Biología y Medicina Experimental (IBYME), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina; Universidad Argentina de la Empresa (UADE), Instituto de Tecnología (INTEC), Ciudad de Buenos Aires, Argentina. Electronic address:
Galectins (Gals), a family of multifunctional glycan-binding proteins, have been traditionally defined as β-galactoside binding lectins. However, certain members of this family have shown selective affinity toward specific glycan structures including human milk oligosaccharides (HMOs) and blood group antigens. In this work, we explored the affinity of human galectins (particularly Gal-1, -3, -4, -7, and -12) toward a panel of oligosaccharides including HMOs and blood group antigens using a complementary approach based on both experimental and computational techniques.
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