Transient expression of chimeric CAT genes injected into early embryos of the domesticated silkworm Bombyx mori.
In order to establish a transient expression system for genes introduced into early embryos of the silkworm, Bombyx mori, we tested various promoters ligated with CAT reporter genes. The embryos into which we injected supercoiled plasmid DNA of pFb(-860/+10)CAT containing the Bombyx fibroin promoter region ligated to the CAT gene showed a reasonably high CAT activity beginning around 30 h after oviposition. This high activity was observed only when the plasmid was injected before termination of the early nuclear cleavage stage, which was about 8 h after oviposition, but not after this stage. This means that the expression of injected DNA is closely related to the presence of cleavage nuclei in early embryos. Promoters originating from insect genes, like the Bombyx sericin-1 gene, Drosophila hsp70 and Drosophila copia LTR, functioned as strong promoters in the embryos. On the contrary, promoters from mammalian virus genes, such as the SV40 early and Moloney murine leukemia virus LTR genes, functioned as weak promoters. Moreover, linearized DNAs showed no or weak activity of expression in embryos. From these results, we conclude that the silkworm embryo transient expression system is a useful tool for studying the mechanism of regulation of insect genes.
Tamura
T
T
National Institute of Sericultural and Entomological Science, Ibaraki.
Kanda
T
T
Takiya
S
S
Okano
K
K
Maekawa
H
H
D000818:Animals; D012831:Bombyx / Q000196:embryology / Q000235:genetics*; D002678:Chimera / Q000235:genetics*; D002970:Cleavage Stage, Ovum; D004247:DNA / Q000235:genetics*; D005349:Fibroins / Q000235:genetics; D005786:Gene Expression Regulation; D005814:Genes, Viral / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic / Q000235:genetics*
D004247:DNA; D005349:Fibroins