[Diagnosis of the sex of bovine embryos using molecular biology].
Thanks to a bovine genomic library enriched with Y chromosome-specific sequences, a probe specific to this chromosome of genera Bos and Bison and repeated on about 2,000 copies in the male genome was isolated. To be compatible with embryo transfer and/or freezing, sex determination has to be done on a 10-20 cell embryo biopsy from a day-7 bovine blastocyst. The in situ hybridization with biotinylated BC 1.2 probe and immunocytochemical revelation permitted the visualization of the hybridization signal on the nucleus of each cell. Because of the numerous steps required, this technique was difficult to apply routinely. For that reason we worked out the polymerase chain reaction technique. From the BC 1.2 sequence, we determined several oligonucleotide primers and probes with the aim of amplifying the Y chromosome-specific sequences. This technique, which is easy to perform and to automate, enabled us to obtain a strong male-specific hybridization signal on genomic DNA and embryo cells.
Laboratoire de Radiobiologie Appliquée, CEA-IPSN-DPS-SPE, Jouy-en-Josas, France.
M
M
Kirszenbaum
C
C
Cotinot
M
M
Leonard
M
M
Vaiman
M
M
Fellous
D000822:Animal Husbandry / Q000379:methods*; D000818:Animals; D001755:Blastocyst; D002417:Cattle / Q000196:embryology* / Q000235:genetics; D004247:DNA / Q000235:genetics; D015342:DNA Probes; D004624:Embryo Transfer; D005260:Female; D015698:Genomic Library; D005865:Gestational Age; D008297:Male; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012732:Sex Determination Analysis; D014998:Y Chromosome
D015342:DNA Probes; D004247:DNA
Diagnostic du sexe des embryons bovins par biologie moléculaire.