Gene integration into bovine sperm genome and its expression in transgenicoffspring

Transgenic bovine sperm were produced by restriction enzyme mediated insert ion (REMI). REMI utilizes lipofection of linearized pEGFP and the correspon ding restriction enzyme for integration into the sperm genomic DNA. The tra nsgenic sperm were used in IVF to produce morula expressing GFP. When trans genic sperm were used for Al in two cows, the resultant calves expressed th e exogenous DNA in their lymphocytes as determined by (a) PCR and RT-PCR; ( b) specific emission of green fluorescence by GFP; and (c) Southern blot an alysis. Data demonstrate that REMI is an efficient method for the productio n of transgenic sperm and corresponding offspring by Al or embryos by IVF. (C) 2000 Wiley-Liss, Inc.