Abstract:

In order to offer doner cells for bovine transgenic cloning by nuclear transfering, fibroblast cells were isolated by attaching tissue explants from bovine fetal. The plasmid containing genes of Caenorhabditis elegans ω-3 fatty acid desaturase Gene was transfected into bovine fetal fibroblast by Lipofectmine, Cell clones were obtained after screening by G418. The recombinant of extrogenous DNA was identified by polymerase chain reaction，the positive colonies were maintained in culture medium containing G418 for 2-3 passages. the transcription of extrogenous DNA was identified by reverse transcription- polymerase chain reaction. Results showed that primary bovine fibroblast cells were isolated from the tissue cultured for 7 day，and the transgenic cells were obtained after G418 selection for 9d. Identification of the transgene in the cell clones was examined by PCR and the exogenous DNA (CMV promoter and Caenorhabditis elegans ω-3 fatty acid desaturase gene) had been integrated into genome. The transgenic cell line could transcript the Caenorhabditis elegans ω-3 fatty acid desaturase DNA efficiently.after passage These results have paved the way to obtain the new transgenic bovine by nuclear transfer in the future.