Patent EP-1649058-B1 (EP)
Inventor: OULMOUDEN AHMAD
IPC: C12Q1/68
Filed: 2004-07-22
Granted: 2010-10-20
Abstract: The use of nucleotide sequences corresponding to the gene SILVER (SI) and to different allelic forms of the gene or corresponding to fragments of the gene or its allelic forms to implement a method of identification of different populations or breeds of ruminant mammifers such as cattle, sheep and goats. Independent claims are included for: (1) The use of nucleotide primers permitting the amplification of the number of copies of the SILVER gene or different allelic forms of the gene as defined to implement a method of identification of different populations or breeds of ruminant mammifers such as cattle, sheep and goats; (2) A nucleotide sequence corresponding to the bovine gene SI, represented by SEQ ID NO: 1 or the following fragments of the gene SI: (a) any fragment from those of about 10-300 nucleotides containing the nucleotides located at positions 82-93 of SEQ ID NO: 1; (b) the fragment SEQ ID NO: 7 of 294 nucleotides delimited by the nucleotides located in positions 9 and 302 of SEQ ID NO: 1; (3) A nucleotide sequence corresponding to the bovine 'si' gene represented by SEQ ID NO: 3 or to the fragments of the gene: (a) among those of about 10-300 nucleotides containing the nucleotides located at positions 82-93 of SEQ ID NO: 3; (b) the fragment of SEQ ID NO: 8 of 8-294 nucleotides delimited by the nucleotides in positions 9 and 302 of SEQ ID NO: 3; (4) A nucleotide sequence corresponding to the bovine gene 'si 1' represented by SEQ ID NO: 5 or the following fragments of the gene: (a) any fragment of about 10-300 nucleotides containing the nucleotides located at positions 82-93 of SEQ ID NO: 5; (b) the fragment SEQ ID NO: 9 of 291 nucleotides delimited by the nucleotides in positions 9 and 299 of SEQ ID NO: 5; (5) A nucleotide sequence corresponding to the claimed primer couple; (6) Primer couples of which each of the primers comprises independently 10-300 nucleotides so chosen that one of the two sequences hybridizes itself with a sequence of about 10-30 nucleotides comprised in the complementary nucleotide sequence delimited by the nucleotides located at positions 1 and about 60 of the sequences SEQ ID NO: 1, 3 and 5, while the other sequence of the couple hybridizes itself with a sequence of about 10-30 nucleotides comprised between the nucleotide located at position 94 and the last of the nucleotides of sequences SEQ ID NO: 1, 3 and 5; (7) A method of identification of populations or breeds of ruminant mammifers such as cattle, sheep and goats carried out starting from a biological sample taken from the animal, especially sperm, embryo, blood, milk, hair, carcase or meat or other products derived from them and permitting the checking and certification that the animal belongs or does not belong to a population or breed comprising: (a) a stage of amplification of a number of copies of the different allelic forms of the SILVER gene, the alleles SI, 'si' or 'si 1' and/or fragments of those allelic forms specific to a particular population or breed and liable to be present in the sample; (b) a stage of detection of the allelic forms or fragments; (8) A kit for the implementation of the method comprising at least the claimed primer couple and, where appropriate, the reagents necessary to implement the amplification reaction.