|
KMID : 1005220090240030225
|
|
Journal of Embryo Transfer
2009 Volume.24 No. 3 p.225 ~ p.230
|
|
|
Sex Ratio Determination by Quantitative Real Time PCR using Amelogenin Gene in Porcine Sperm
|
|
Hwang You-Jin
Kim Dae-Young
Park Choon-Keun
Bae Mun-Sook
Yang Jae-Hun
Kim Bo-Kyung
Kim Sang-Ok
Lee Eun-Soo
Choi Sun-Gyu
Kwon Ye-Ri
Seo Min-Hae
|
|
|
Abstract
|
|
|
|
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.
|
|
|
KEYWORD
|
|
|
amelogenin (AMEL) gene, sex determination, quantitative real time PCR, boar sperm
|
|
|
FullTexts / Linksout information
|
|
|
|
|
Listed journal information
|
|
|