Expression of a mouse metallothionein-Escherichia coli beta-galactosidase fusion gene (MT-beta gal) in early mouse embryos.
We have microinjected DNA containing the inducible mouse metallothionein-I (MT-I) promoter, coupled to the structural gene for Escherichia coli beta-galactosidase (lacZ), into the pronuclei of one-cell mouse embryos. A qualitative histochemical assay, with 5-bromo-4-chloro-3-indolyl beta-D-galactopyranoside (X-Gal) as a substrate, was used to detect expression of lacZ at several preimplantation stages. We observed staining indicative of exogenous beta-galactosidase activity in 5-17% of DNA-injected embryos assayed at preimplantation stages after 16-24 h treatment with ZnSO4. Thus, lacZ can be used as an indicator gene for promoter function during early mouse embryogenesis, and the incorporation of the MT-I promoter into fusion genes can be a useful means of controlling the expression of exogenous genes in preimplantation mouse embryos.
Stevens
M E
ME
Laboratory of Radiobiology and Environmental Health, University of California, San Francisco 94143.
Meneses
J J
JJ
Pedersen
R A
RA
D000818:Animals; D001755:Blastocyst / Q000378:metabolism; D004274:DNA, Recombinant / Q000378:metabolism; D004622:Embryo, Mammalian / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005260:Female; D005696:Galactosidases / Q000235:genetics*; D005796:Genes; D005798:Genes, Bacterial; D008668:Metallothionein / Q000096:biosynthesis / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D008845:Microinjections; D009028:Morula / Q000378:metabolism; D009030:Mosaicism; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D015183:Restriction Mapping; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics*
D004274:DNA, Recombinant; D011993:Recombinant Fusion Proteins; D008668:Metallothionein; D005696:Galactosidases; D001616:beta-Galactosidase