한국수정란이식학회지 (Journal of Embryo Transfer)

  • 제24권3호
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  • Pages.225-230
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  • 2009
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  • 2508-755X(pISSN)
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  • 2288-0178(eISSN)
한국수정란이식학회 (The Korean Society of Embryo Transfer)
권호 표지

Sex Ratio Determination by Quantitative Real Time PCR using Amelogenin Gene in Porcine Sperm

  • Hwang, You-Jin (Division of Biological Science, Gachon University of Medicine and Science) ;
  • Bae, Mun-Sook (Division of Biological Science, Gachon University of Medicine and Science) ;
  • Yang, Jae-Hun (Division of Biological Science, Gachon University of Medicine and Science) ;
  • Kim, Bo-Kyoung (Division of Biological Science, Gachon University of Medicine and Science) ;
  • Kim, Sang-Ok (Division of Biological Science, Gachon University of Medicine and Science) ;
  • Lee, Eun-Soo (Division of Biological Science, Gachon University of Medicine and Science) ;
  • Choi, Sun-Gyu (Division of Biological Science, Gachon University of Medicine and Science) ;
  • Kwon, Ye-Ri (Division of Biological Science, Gachon University of Medicine and Science) ;
  • Seo, Min-Hae (Division of Biological Science, Gachon University of Medicine and Science) ;
  • Park, Choon-Keun (College of Animal Life Science, Kangwon National University) ;
  • Kim, Dae-Young (Division of Biological Science, Gachon University of Medicine and Science)
  • 발행 : 2009.09.30
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초록

Sex-sorting of sperm is an assisted reproductive technology (ART) used by the livestock industry for the mass production of animals of a desired sex. The standard method for sorting sperm is the detection of DNA content differences between X and Y chromosome-bearing sperm by flow cytometry. However, this method has variable efficiency and therefore requires verification by a second method. We have developed a sex determination method based on quantitative real-time polymerase chain reaction (qPCR) of the porcine amelogenin (AMEL) gene. The AMEL gene is present on both the X and the Y chromosome, but the length and sequence of its noncoding regions differ between the X and Y chromosomes. By measuring the threshold cycle (Ct) of qPCR, we were able to calculate the relative frequency of X chromosome. Two sets of AMEL primers were used in these studies. One set (AME) targeted AMEL gene sequences present in both X and Y chromosome, but produced PCR products of different lengths for each chromosome. The other set (AXR) bound to AMEL gene sequences present on the X chromosome but absent esholthe Y-chromosome. Relative product levels were calculated by normalizing the AXR fluorescence to the AME fluorescence. The AMEL method accurately predicted the sex ratios of boar sperm, demonstrating that it has potential value as a sex determination method.

키워드

참고문헌

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