Authors: I A Burkov I A Serova N R Battulin A V Smirnov I V Babkin L E Andreeva G A Dvoryanchikov O L Serov
Publish Date: 2013/02/23
Volume: 22, Issue: 5, Pages: 949-964
Abstract
Expression of the human granulocyte–macrophage colonystimulating factor hGMCSF gene under the control of the 5′regulatory sequence of the goat alphaS1casein 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 tissuesspecific expression of the hGMCSF gene in the mammary gland only and no signs of cell mosaicism The concentration of hGMCSF in the milk of transgenic females varied from 19 to 14 μg/ml One line presented hGMCSF in the blood serum indicating ectopic expression The values of secretion of hGMCSF in milk of 6 transgenic lines carrying the transgene with MAR varied from 005 to 07 μg/ml and two of these did not express hGMCSF Three of the four examined animals from lines of this group showed ectopic expression of the hGMCSF gene as determined by RTPCR and immunofluorescence analyses as well as the presence of hGMCSF in the blood serum Mosaic expression of the hGMCSF 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 hGMCSF gene expression was not observedThe authors express gratitude to Dr Nelly Khaidarova Institute of Molecular Genetics Academy of Sciences of Russia Moscow Russia for help in preparation of recombinant DNA and Dr Irina Gileva for providing us the recombinant human GMCSF derived from E coli This study was financially supported by Grant5626 from Program of Fundamental Research RAN
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