Inducible expression of a foreign gene inserted into the human cytomegalovirus genome.
We previously described the insertion of a foreign gene into a non-essential region of human cytomegalovirus (HCMV) by homologous recombination. Here we report insertion of the Escherichia coli lacZ gene downstream of the mouse metallothionein promoter into the HindIII-O region of HCMV by replacement-type recombination. Expression of the lacZ gene in the recombinant was independent of viral growth, but dependent on induction by heavy metals. Of several metals tested for beta-galactosidase induction and also for their toxicity to HEL cells, Zn was found to be the most suitable for use as an inducer. In HEL cells infected with the recombinant in the presence of 50 microsM-Zn, beta-galactosidase activity was maximal 3 days after infection, and reached levels 27 times higher than the value obtained in the absence of Zn.
Takekoshi
M
M
Division of Molecular Life Science, Tokai University School of Medicine, Isehara, Japan.
Maeda-Takekoshi
F
F
Ihara
S
S
Sakuma
S
S
Watanabe
Y
Y
D003587:Cytomegalovirus / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D015967:Gene Expression Regulation, Viral / Q000187:drug effects / Q000235:genetics*; D005798:Genes, Bacterial / Q000235:genetics; D006801:Humans; D008668:Metallothionein / Q000235:genetics*; D008670:Metals / Q000494:pharmacology; D011401:Promoter Regions, Genetic / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000187:drug effects; D011995:Recombination, Genetic / Q000235:genetics*; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000187:drug effects
D008670:Metals; D011993:Recombinant Fusion Proteins; D008668:Metallothionein; D001616:beta-Galactosidase