Isolation and characterization of the mouse metallothionein-I gene.

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Abstract
Double-stranded c[complementary]DNA was synthesized from a mouse liver mRNA fraction enriched for metallothionein mRNA activity, ligated to restriction site linkers, inserted into pBR322, and used to transform Escherichia coli .chi. 1776. The sequence of the largest plasmid containing DNA that hybridized to metallothionein mRNA contained a 380 base pair insert that includes the entire coding region and 3'' untranslated region of metallothionein-I. The metallothionein-I insert was nick translated and used to screen a mouse myeloma and a mouse embryo DNA library in bacteriophage .lambda.. A metallothionein-I genomic clone containing 13-15 kbase pairs of mouse DNA was isolated from each library. Both contain a 3.8 kbase pair EcoRI fragment that hybridizes to the metallothionein-I probe. The location, size and orientation of the metallothionein-I gene within the 3.8 kbase pair fragment were determined by heteroduplex and restriction mapping. The gene spans 1.1 kbase pairs and contains at least 2 introns.