Science · 1993 · 118 citations · 31 references
Life SciencesGeneticsMolecular BiologyGene ExpressionGenetic EngineeringTransgenic Animal ModelsEmbryonic Stem CellsTransgene ConstructionTransgenesis StrategiesTransgenic Animal TechnologyTransgenic Organism DevelopmentYeast Artificial ChromosomesCol1a1 MutationsEmbryonic Stem Cell BiologyEmbryonic Stem Cell-based TransgenesisTransgene ExpressionTransgene Expression AnalysisTransgenic Cell LinesTransgenic MiceTransgenic Mouse Models
Molecular complementation of mutant phenotypes by transgenic technology is a potentially important tool for gene identification. A technology was developed that allows the transfer of a physically intact yeast artificial chromosome (YAC) into the germ line of the mouse. A purified 150-kilobase YAC encompassing the murine gene Col1a1 was efficiently introduced into embryonic stem (ES) cells via lipofection. Chimeric founder mice were derived from two transfected ES cell clones. These chimeras transmitted the full length transgene through the germ line, generating two transgenic mouse strains. Transgene expression was visualized as nascent transcripts in interphase nuclei and quantitated by ribonuclease protection analysis. Both assays indicated that the transgene was expressed at levels comparable to the endogenous collagen gene.
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Douglas A. Melton, Paul A. Krieg, Michael R. Rebagliati et al. · Nucleic Acids Research · 1984 · 6.4K citations · Full text
A second-generation linkage map of the human genome
Jean Weissenbach, Gàbor Gyapay, Colette Dib et al. · Nature · 1992 · 1.7K citations