Expression of the human granulocyte–macrophage colony stimulating factor (hGM-CSF) gene under control of the 5′-regulatory sequence of the goat alpha-S1-casein gene with and without a MAR element in transgenic mice
| Autor: | Gennady Dvoryanchikov, I. V. Babkin, Oleg L. Serov, Nariman Battulin, Alexander V. Smirnov, Serova Ia, Lyudmila Andreeva, I. A. Burkov |
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| Rok vydání: | 2013 |
| Předmět: |
Male
Genetically modified mouse Transgene Fluorescent Antibody Technique Enzyme-Linked Immunosorbent Assay Mice Transgenic Regulatory Sequences Nucleic Acid Biology Polymerase Chain Reaction Histones Mice Mammary Glands Animal Blood serum Gene expression Genetics Animals Humans Cloning Molecular Enhancer Gene DNA Primers Reverse Transcriptase Polymerase Chain Reaction Goats Caseins Granulocyte-Macrophage Colony-Stimulating Factor Matrix Attachment Regions Molecular biology Regulatory sequence Drosophila Female Animal Science and Zoology Ectopic expression Agronomy and Crop Science Plasmids Biotechnology |
| Zdroj: | Transgenic Research. 22:949-964 |
| ISSN: | 1573-9368 0962-8819 |
| DOI: | 10.1007/s11248-013-9697-4 |
| Popis: | Expression of the human granulocyte-macrophage colony-stimulating factor (hGM-CSF) gene under the control of the 5'-regulatory sequence of the goat alpha-S1-casein gene with and without a matrix attachment region (MAR) element from the Drosophila histone 1 gene was studied in four and eight transgenic mouse lines, respectively. Of the four transgenic lines carrying the transgene without MAR, three had correct tissues-specific expression of the hGM-CSF gene in the mammary gland only and no signs of cell mosaicism. The concentration of hGM-CSF in the milk of transgenic females varied from 1.9 to 14 μg/ml. One line presented hGM-CSF in the blood serum, indicating ectopic expression. The values of secretion of hGM-CSF in milk of 6 transgenic lines carrying the transgene with MAR varied from 0.05 to 0.7 μg/ml, and two of these did not express hGM-CSF. Three of the four examined animals from lines of this group showed ectopic expression of the hGM-CSF gene, as determined by RT-PCR and immunofluorescence analyses, as well as the presence of hGM-CSF in the blood serum. Mosaic expression of the hGM-CSF gene in mammary epithelial cells was specific to all examined transgenic mice carrying the transgene with MAR but was never observed in the transgenic mice without MAR. The mosaic expression was not dependent on transgene copy number. Thus, the expected "protective or enhancer effect" from the MAR element on the hGM-CSF gene expression was not observed. |
| Databáze: | OpenAIRE |
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