Effects of Germ Line Transmission in Cloned Calves Derived from In Vitro-Transfected Somatic Cells

摘要

In vitro trartsfecdon of cultured cells combined with nuclear trailsfer currently is the moat eriective procedure to produce transgenic livestock. In the present study,bovine primary fetal flbroblasts were transfected with a green fluorescent protein (GFP)reporter transgene and used as nuclear donor cells in ooevte reconstructions．To examine the role of host cytoplasm on transgene expression and developmenbl outcome．GFP-expressing fibroblasts were fused to oocytes reconstrucled either metaphase or telophase activation,and PCR technology was also employed．The results showed that GFP became detectahie at the 8-to 16-cell stage， approximately 80 h after reconstruction，and remained positive at all later stages．Embrvonic development to the blastocvst stage was not significantly dlfferent among metaphase and telophase groups．Therefore，GFP transgene technology can be used to select embryoes derived frum transgenic animals．