家畜の性決定遺伝子(SRY)保存領域のPCR法による増幅と塩基配列<sup>†</sup>の種差

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  • Amplification and Sequence Analysis of SRY (sex-determining region Y) Conserved Region of Domestic Animals Using Polymerase Chain Reaction
  • カチク ノ セイ ケッテイ イデンシ SRY ホゾン リョウイキ ノ PCRホ

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Previous studies have shown that SRY (sex-determining region of Y) was sufficient to direct the formation of testis, and SRY has been found on the Y chromosomes of all other eutherian mammals tested by the Southern blot analysis. This study was conducted to detect the SRY fragments of domestic animal using the Polymerase Chain Reaction (PCR) with the primers made from the conserved sequence of human, mouse and rabbit. In addition, the sequences of the amplified fragments were determined by direct sequencing and were compared with the SRY sequences of human, mouse and rabbit. We examined each male and female of human, cattle, goat, sheep, pig, deer and mouse for PCR. The synthetic oligonucleotides were used as primers; primer 1 (sense primer), 5' AAGCGACCCATGAATGCATTCATGGTGTGGT 3' (31mer): primer 2 (antisence primer), GAGGTCGATACTTATAGTTCGGGTATTTCTCTCTGTG 3' (37mer). PCR reaction was performed with 1 unit of Tth DNA polymerase for 35 cycles of amplification (denaturation at93°C for 1 min, annealing at 60°C for 1.5min, extention at 72°C for 1min). After amplification, the male specific DNA band which was similar in size to that of the human and the mouse was detected in each species. Subsequently, we determined the sequences of these fragments. It was found that there were considerable differences among the DNA sequences of the SRY conserved region of domestic animals, but these amino acid sequences were well-conserved for their function. Therefore, it seems that the region we examined coded the protein which play an important part as the sex-determining gene.

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  • 日本畜産学会報

    日本畜産学会報 63 (10), 1059-1065, 1992

    公益社団法人 日本畜産学会

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