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A yeast artificial chromosome covering the tyrosinase gene confers copy number-dependent expression in transgenic mice

Item Type:Article
Title:A yeast artificial chromosome covering the tyrosinase gene confers copy number-dependent expression in transgenic mice
Creators Name:Schedl, A. and Montoliu, L. and Kelsey, G. and Schutz, G.
Abstract:Expression of transgenes in mice often fails to follow the normal temporal and spatial pattern and to reach the same level as the endogenous copies. Only in exceptional cases has position-independent and copy number-dependent expression been reproduced. The size constraint of standard constructs may prevent the inclusion of important remote regulatory elements. Yeast artificial chromosomes (YACs) provide a means of cloning large DNA fragments and the transfer of YAC DNA into somatic cells has been reported. We have previously produced transgenic mice carrying a 35 kilobase YAC construct. Here we report the transfer of a 250 kilobase YAC covering the mouse tyrosinase gene into mice by pronuclear injection of gel-purified YAC DNA. The YAC was inserted into the mouse genome without major rearrangements and expression of the YAC-borne tyrosinase gene resulted in complete rescue of the albino phenotype of the recipient mice. Expression from the transgene reached levels comparable to that of the endogenous gene and showed copy number dependence and position independence.
Keywords:Base Sequence, Fungal Chromosomes, Genetic Crosses, DNA Probes, Exons, Genetic Techniques, Hair Color, Molecular Sequence Data, Monophenol Monooxygenase, Oligodeoxyribonucleotides, Oocytes, Polymerase Chain Reaction, Restriction Mapping, Saccharomyces cerevisiae, Genetic Transcription, Animals, Mice
Source:Nature
ISSN:0028-0836
Publisher:Nature Publishing Group (U.K.)
Volume:362
Number:6417
Page Range:258-261
Date:18 March 1993
Official Publication:https://doi.org/10.1038/362258a0
PubMed:View item in PubMed

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