Process for sexing cow embryos
First Claim
1. An oligonucleotide primer set for bovine embryo sexing wherein said oligonucleotide primer set comprises a primer pair which primer pair consists of individual oligonucleotide primers each of which is able to hybridize specifically and simultaneously to an intron 5 sequence of the bovine amelogenin gene, which gene is located on both the bovine X and Y chromosomes, wherein said intron 5 sequence of the bovine amelogenin gene is selected from the group consisting of:
- SEQ ID NO;
21 (the nucleotide sequence of intron 5 of the bovine amelogenin gene located on the bovine X chromosome), that sequence fully complementary to SEQ ID NO;
21, SEQ ID NO;
22 (the nucleotide sequence of intron 5 of the bovine amelogenin gene located on the bovine Y chromosome), and that sequence fully complementary to SEQ ID NO;
22.
1 Assignment
0 Petitions
Accused Products
Abstract
A rapid, highly reproducible and sensitive technique has been successfully developed for sexing the cow embryos, by method of polymerase chain reaction (PCR) against the amelogenin (bAML) genes located on both X- and Y-chromosomes of the Holstein dairy cattle. Results from DNA sequence analysis showed that there was only 45% homology between the intron 5 of AMLX and AMLY genes. Based on these sequences a pair of sex-specific primers, pbAML5XY(+) and pbAML5XY (-),were designed allowing to amplify a single fragment of 476-bp from the female cattle and two fragments of 476-bp and 341-bp from the male ones, respectively. The most important feature is that the precise sensitivity of sex-determination was confirmed to be reached as minimum template as trace amount of genomic DNA content in either a single lymphocyte or a single blastomere isolated from cow embryo at day-6 to day-7. Moreover, neither those of complicated procedures for purifying the DNA prior the PCR nor any extra pair of primers for serving as internal control is thought to be essential and the sex-determination of over hundred embryos can be completed at once within 4hrs.
-
Citations
7 Claims
-
1. An oligonucleotide primer set for bovine embryo sexing wherein said oligonucleotide primer set comprises a primer pair which primer pair consists of individual oligonucleotide primers each of which is able to hybridize specifically and simultaneously to an intron 5 sequence of the bovine amelogenin gene, which gene is located on both the bovine X and Y chromosomes, wherein said intron 5 sequence of the bovine amelogenin gene is selected from the group consisting of:
- SEQ ID NO;
21 (the nucleotide sequence of intron 5 of the bovine amelogenin gene located on the bovine X chromosome), that sequence fully complementary to SEQ ID NO;
21, SEQ ID NO;
22 (the nucleotide sequence of intron 5 of the bovine amelogenin gene located on the bovine Y chromosome), and that sequence fully complementary to SEQ ID NO;
22. - View Dependent Claims (4, 7)
- SEQ ID NO;
-
2. A primer pair, for sex determination of cow embryonic cells, which comprises a DNA sequence selected from the group consisting of:
-
5'"'"'-AAATTCTCTCACAGTCCAAG-3'"'"' (SEQ ID NO;
16); and5'"'"'-CAACAGGTAATTTTCCTTTAG-3'"'"' (SEQ ID NO;
19). - View Dependent Claims (3, 5)
-
-
6. A compound which is a oligonucleotide selected from the group consisting of SEQ ID NO:
- 14, SEQ ID NO;
16, SEQ ID NO;
19, SEQ ID NO;
21 and SEQ ID NO;
22.
- 14, SEQ ID NO;
Specification