Abstract：Abstract: In order to prepare the nuclear donor cells for bovine transgenic cloning, bovine fibroblast cells were isolated by attaching tissue explants from ear skin of a bovine. Then the cells were purified and the cell growth curve was plotted. The purified plasmid containing two genes of hLYZ and EGFP was transfected by lipofectmine 2000 into bovine fibroblast. The cells were checked 24h hours after transfection under fluorescence microscopy for EGFP expression, and G418 selection (500μg/ml) was applied since then. After 2 weeks, positive colonies were maintained in culture medium containing 250μg/ml G418 for 2 - 3 passages. A small part of the positive cells were analyzed by PCR for gene integration. PCR detection demonstrated that the foreign gene was integrated into the genome. The results indicated that the isolated bovine fibroblast cells might be competent for transgenic cloning.
