Sex determination of equine somatic and germ cells by PCR amplification based on the sequence polymorphism of X-and Y-chromosomal amelogenin genes
1999
Fukushima, Y. (Tochigi Prefectural Police Headquarters, Utsunomiya (Japan)) | Mukoyama, H. | Sato, F. | Hasegawa, T. | Ishida, N. | Muramatsu, S.
Two distinct transcripts for equine X-and Y-chromosomal amelogenin genes were cloned from male and female fetal ameloblast cDNA libraries. The complete nucleotide sequence of equine X-chromosomal amelogenin cDNA was 696-base-pairs (bp) that encodes a protein with 192 amino acid residues. The deletion of 24 bp was found in the fifth exon of equine Y-chromosomal amelogenin gene. The genomic DNAs of equine male and female were amplified by the polymerase chain reaction using a pair of nucleotide primers from an X/Y-homologous region of amelogenin gene. The PCR product from mare genomic DNA revealed a single band (184 bp) corresponding to the X-chromosome by 4 % agargel electrophoresis. And the PCR products from stallion and gelding genomic DNA showed three bands which were two distinct bands (184 bp and 160 bp) with X-and Y-chromosome and an additional band (to 200 bp) of heteroduplex. This PCR polymorphism of X-and Y-chromosomal amelogenin genes enabled sex determination of single s
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