Distal regulatory elements from the mouse metallothionein locus stimulate gene expression in transgenic mice.
DNA regions of 10 and 7 kb that flank the mouse metallothionein II (MT-II) and MT-I genes, respectively, were combined with a minimally marked MT-I (MT-I*) gene and tested in transgenic mice. This construct resulted in (i) position-independent expression of MT-I* mRNA and copy number-dependent expression, (ii) levels of hepatic MT-I mRNA per cell per transgene that were about half that derived from endogenous MT-I genes, (iii) appropriate regulation by metals and hormones, and (iv) tissue distribution of transgene mRNA that resembled that of endogenous MT-I mRNA. These features were not observed when MT-I* was tested without the flanking regions. These MT-I flanking sequences also improved the expression of rat growth hormone reporter genes, with or without introns, that were under the control of the MT-I promoter. Moreover, they enhanced expression from two of four heterologous promoters/enhancers that were tested. Deletion analysis indicated that regions known to have DNase I-hypersensitive sites were necessary but not sufficient for high-level expression. These data suggest that the DNA regions flanking the mouse MT-I and MT-II genes have functions like the locus control regions described for other genes.
Palmiter
R D
RD
Department of Biochemistry, Howard Hughes Medical Institute, University of Washington SL-15, Seattle 98195.
Sandgren
E P
EP
Koeller
D M
DM
Brinster
R L
RL
D000818:Animals; D001483:Base Sequence; D003850:Deoxyribonuclease I / Q000494:pharmacology; D004742:Enhancer Elements, Genetic; D005260:Female; D005786:Gene Expression Regulation; D005796:Genes; D008297:Male; D008668:Metallothionein / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid; D015032:Zinc / Q000494:pharmacology
D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D008668:Metallothionein; D003850:Deoxyribonuclease I; D015032:Zinc