Sex Determination of Equine Somatic and Germ Cells by PCR Amplification Based on the Sequence Polymorphism of X- and Y-Chromosomal Amelogenin Genes

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  • アメロゲニン遺伝子の多型性を指標としたウマの体細胞および生殖細胞の性別判定法
  • アメロゲニン イデンシ ノ タケイセイ オ シヒョウ ト シタ ウマ ノ タイサイボウ オヨビ セイショク サイボウ ノ セイベツ ハンテイホウ

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Abstract

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 24bp 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 (184bp) corresponding to the X-chromosome by 4% agar gel electrophoresis. And the PCR products from stallion and gelding genomic DNA showed three bands which were two distinct bands (184bp and 160bp) with X-and Y-chromosome and an additional band (-200bp) of heteroduplex. This PCR polymorphism of X-and Y-chromosomal amelogenin genes enabled sex determination of single sperm as well as equine culture cells.

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