Gene integration into bovine sperm genome and its expression in transgenic offspring.
Transgenic bovine sperm were produced by restriction enzyme mediated insertion (REMI). REMI utilizes lipofection of linearized pEGFP and the corresponding restriction enzyme for integration into the sperm genomic DNA. The transgenic sperm were used in IVF to produce morula expressing GFP. When transgenic sperm were used for AI in two cows, the resultant calves expressed the 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 analysis. Data demonstrate that REMI is an efficient method for the production of transgenic sperm and corresponding offspring by AI or embryos by IVF.
Shemesh
M
M
Department of Hormone Research, Kimron Veterinary Institute, Bet Dagan, Israel. mshem@vs.moag.gov.il
Gurevich
M
M
Harel-Markowitz
E
E
Benvenisti
L
L
Shore
L S
LS
Stram
Y
Y
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics* / Q000378:metabolism; D000831:Animals, Newborn; D015139:Blotting, Southern; D002417:Cattle; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004622:Embryo, Mammalian / Q000378:metabolism; D005307:Fertilization in Vitro; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins; D066298:In Vitro Techniques; D007315:Insemination, Artificial; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013094:Spermatozoa; D014162:Transfection
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D004247:DNA; D004262:DNA Restriction Enzymes