The effects of one strain Agrobacterium tumefaciens LBA 4404/pBI 121, conditions of co-cultivation and growth regulators on regeneration were studied and trials of transformation were performed on winter oilseed rape Brassica napus L. cv. Lirajet, Valesca and Bor. Strong shoots regeneration was observed after co-cultivation without acetosyringone. Among antibiotics that were used in the experiments, kanamycin A and geneticin hampered whereas carbenicillin and rifampicin stimulated shoots regeneration. The presence of nptII gene was confirmed by PCR and K1, K2 primers which generate a fragment of 732 bp in length.