• Journal of Life Science
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• v.17 no.3 s.83
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• pp.402-406
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• 2007

This study was performed to develop the molecular marker for sex determination of hare (Lepus coreanus) distributed in Korea which focused on sexual dimorphism between X and Y chromosomal homologous genes, zinc finger-X (ZFX) and -Y (ZFY). The intron 7 regions of ZFX and ZFY genes exhibited differential amplification patterns between male and female hares. The lengths of intron 7 region of ZFX and ZFY genes were 538 and 233-bp, respectively. Especially, the ZFX intron 7 contained a repetitive sequence identified as member of RNA-mediated transposable elements which was similar to CSINE2 commonly found in the rabbit genome. However, it was not present in intron 7 of ZFY gene. The molecular sex typing by polymerase chain reaction (PCR) was also carried out to determine the sex of hare based on difference in lengths between the intron 7 regions of ZFX and ZFY genes. All DNA samples tested had common band amplified from ZFX. However, the male hare DNAs had two distinct bands which amplified from ZFX and ZFY genes, respectively. The results from ZFX-ZFY PCR sex typing were identical to those from phenotypic investigation and from amplification patterns using male-specific sex determining region Y (SRY) gene as well. Finally, this study suggested that the sexual dimorphism between intron 7 regions of ZFX and ZFY could be useful genetic marker to determine sex of hare.

• Journal of Animal Science and Technology
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• v.49 no.1
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• pp.1-8
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• 2007

This study was focused on discriminating the molecular sexes of red deer and elk by duplex polymerase chain reaction(PCR) using two primer sets. Sex differentiation of mammals is primarily dependent on the presence or absence of sex determining region Y(SRY) gene encoded on Y chromosome which plays a key role for male development. Zinc finger X-Y(ZFX-ZFY) gene, one of X-Y homology gene group was found on X- and Y- chromosomes, respectively. At first, the nucleotide sequences were characterized for the intron 9 flanking region of ZFX-ZFY genes. The intron 9 of ZFX and ZFY is 529-bp and 665-bp in length, respectively. A transposable element sequence similar to bovine SINE element Bov-tA was detected only in ZFY gene of Cervidae. Sexing analysis was conducted by duplex PCR assay for amplification of SRY and ZFX-ZFY genes. Two differentially amplified patterns were found: one for females has a common band amplified only from ZFX as a template, and another for males had three bands(a common ZFX and two male-specific ZFY and SRY). On the separate tests using each gene, the results was identical to those from duplex PCR assay. Moreover, the results from PCR assays provide also identical information to phenotypic investigation of individuals of red deer, elk as well as their hybridized progenies collected from two isolated farms. These results suggest that it may be a rapid and precise method for determining the sexes by duplex PCR amplification using Y-chromosome specific SRY and X- and Y- homologous ZFX-ZFY genes showing sexual dimorphism in red deer and elk without any other controls.

• Food Science of Animal Resources
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• v.27 no.3
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• pp.351-356
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• 2007

The objective of this study was to develop a rapid and reliable method for the sex determination of beef using the PCR(polymerase chain reaction) technique. We have used two bovine sex determining genes, SRY and ZFY, on the Y-chromosome to identify the sex of Hanwoo and Holstein beet. We attempted to amplify 1,348 bp and 979 bp fragments from male and female genomic DNA corresponding to the SRY and ZFY genes, respectively, using male specific primers. The amplified PCR products were separated by electrophoresis in a 1.5% agarose gel to detect a male specific DNA band. When DNA from male beef was amplified with primers specific for the SRY gene, a DNA band of 1,348 bp was present in all of the male samples, but absent from all of the female samples. Also, when DNA from male beef was amplified with primers specific for the ZFY gene, a DNA band of 979 bp was observed in all of the male samples, but absent from all female samples. In conclusion, the bovine SRY and ZFY genes are typically found only in male beef. For the practical application of this method for the sexing of commercial beef at the processing and marketing stages after slaughter. a total of 350 beef samples collected randomly from local markets were analyzed for sex determination. The proportions of male and female samples were 252 (72%) and 98 (28%), respectively. Therefore. the SRY and ZFY genes. which are specific for the Y-chromosome, may be useful sex-diagnostic DNA markers to distinguish male meat from female meat.

• Korean Journal of Poultry Science
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• v.20 no.4
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• pp.177-188
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• 1993

This study was performed to find out the reasonable sexing methods In the chicken, obtain the basic information for the mechanisms related to chicken sexual differentiation and identify the genes which known to involved in chicken sex differentiation. The chromosome analysis of chicken embryonic fibroblast was a simple method to determine sex of chicken by means of Z and W chromosome identification. The bands of female chicken genomic DNA digested with Xho Ⅰ and Eco RI restriction endonuclease showed to be useful in direct sex determination and these repetitive sequences of Xho Ⅰ and Eco RI families were proposed to be very homologous in their sequences by colony hybridization analysis. Seven of 150 random primers were selected to amplify the W chromosome-specific band by using arbitrary primed PCR and three of them were useful to identify the sex of chicken. To identify the sex differentiation genes in the chicken, PCR for the amplification of ZFY and SRY sequences was performed. ZFY and SRY sequences were amplified successfully in the chicken genome, implying that chicken genome might have the sex-related conserved sequences similar to mammalian ones. The PCR products of ZFY amplification were the same in both sexes, suggesting that these sequences may be located on autosome or Z chromosome. The profile of PCR amplification for SRY sequences showed variation between sexes, but this result was not enough to specify whether the SRY gene in chicken is on the autosome or sex chromosome.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2005.05a
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• pp.191-194
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• 2005

본 연구는 polymerase chain reaction(PCR)기법을 이용하여 SRY 및 ZFY의 성 결정 유전자의 특정 염기서열을 포함하는 primer를 이용하여 Y-염색체 특이적인 쇠고기 성판별 기술을 개발하기 위해 수행하였다. 성 결정 유전자의 영역에 특정 염기서열을 포함하는 primer를 설계 합성하고 이들 primer를 이용하여 PCR 증폭을 실시한 다음, 각각의 증폭산물을 1.5% agarose gel에 전기영동 하여 웅성 특이적 DNA band의 증폭여부를 확인하였다. SRY 유전자에서 웅성개체 쇠고기는 1,348 bp 크기의 단편을 가진 DNA band가 검출되었으나, 자성개체의 경우 DNA band가 전혀 검출되지 않은 것을 확인 할 수 있었다. 또한, ZFY 유전자에서 웅성개체의 쇠고기는 979 bp 크기의 단편을 가진 DNA band가 모두 검출되었으나, 자성개체의 쇠고기에서는 역시 DNA band가 전혀 검출되지 않았다. 즉,SRY 및 ZFY 유전자는 모두 수소에서 유래한 쇠고기에서 웅성 특이적인 DNA band가 정확히 검출된 반면 암소에서 유래한 쇠고기에서는 웅성 특이적 DNA band가 전혀 검출되지 않았다. 따라서, 본 연구에서 개발한 SRY 또는 ZFY의 웅성 특이적 성 결정유전자를 이용하는 쇠고기 성 감별기술은 시중에서 유통 판매되고 있는 쇠고기의 암수 성감별을 위한 유용한 DNA marker(DNA 표지인자)로 활용할 수 있을 것이다.

• Proceedings of the Korean Society of Life Science Conference
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• 2007.10a
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• pp.52.1-52.1
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• 2007

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• Asian-Australasian Journal of Animal Sciences
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• v.16 no.5
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• pp.650-654
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• 2003

The accuracy of Polymerase chain reaction (PCR) assay in sexing of sheep embryos was assessed in this study. A total of 174 ovine embryos produced in vitro at different stages of development (2, 4-8 cell stages, morula and blastocyst) were sexed. The universal primers (P1-5EZ and P2-3EZ) used in this assay amplified ZFY/ZFX-specific sequences and yielded a 445 bp fragment in both sexes. Restriction enzyme analysis of ZFY/ZFX-amplified fragments with Sac I exhibited polymorphism between sexes, three and two fragments in males and in females, respectively. For verification of accuracy, blood samples of known sex were utilized as positive controls in each test. The mean percentages of sex identification by this method at 2 cell, 4-8 cell, morula and blastocyst were $73.00{\pm}5.72$, $89.77{\pm}3.79$, $3.33{\pm}8.08$ and $79.6{\pm}9.09$, espectively with the over all male to female ratio of 1:0.87. It is concluded that the ZFY/ZFX based method is highly reliable for the sexing of sheep embryos.

• Journal of Embryo Transfer
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• v.31 no.1
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• pp.61-64
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• 2016

We developed a polymerase chain reaction (PCR)-based molecular method for sexing and identification using sexual dimorphism between the Zinc Finger-X and -Y (ZFX-ZFY) gene and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for mitochondrial DNA (mtDNA) cytochrome B (CYTB) gene in meat pieces and commercial sausages from animals of different origins. Sexual dimorphism based on the presence or absence of SINE-like sequence between ZFX and ZFY genes showed distinguishable band patterns between male and female DNA samples and were easily detected by PCR analyses. Male DNA had two PCR products appearing as distinct two bands (ZFX and ZFY), and female DNA had a single band (ZFX). Molecular identification was carried out using PCR-RFLP of CYTB gene, and showed clear species classification results. The results yielded identical information on the sexes and the species of the meat samples collected from providers without any records. The analyses for DNA isolated from commercial sausage showed that pig was the major source but several sausages originated from chicken and Atlantic cod. Applying this PCR-based molecular method was useful and yielded clear sex information and identified the species of various tissue samples originating from livestock.

• Journal of Genetic Medicine
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• v.2 no.1
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• pp.11-15
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• 1998

This paper reports 3 cases with 46,XX sex reversed male. Three 46,XX hypogonadal subjects showed complete sex reversal and had normal phallus and azoospermia. We studied them under clinical, cytogenetic and molecular aspects to find out the origin of the sex reversal. Patients had markedly elevated serum follicle-stimulating hormone (FSH) and lutenizing hormone (LH) and decreased or normal range of serum testosterone. The testicular volumes were small (3-8ml). Testicular biopsy showed Leydig cell hyperplasia and atrophy of seminiferous tubules. We obtained the results of normal 46,XX, and the presence of Y chromosome mosaicism was ruled out through XY dual fluorescent in situ hybridization (FISH). By using polymerase chain reaction (PCR), we amplified short arm (SRY, PABY, ZFY and DYS14), centromere (DYZ3), and heterochromatin (DYZ1) region of the Y chromosome. PCR amplification of DNA from these patients showed the presence of the sex-determining region of the Y chromosome (SRY) but didn't show the centromere and heterochromatin region sequence. The SRY gene was detected in all the three patients. Amplification patterns of the other regions were different in these patients; one had four amplified loci (PABY+, SRY+, ZFY+, DYS14+), another had two loci (SRY+, ZFY+) and the other had two loci (PABY+, SRY+). We have found that each patient's translocation elements had different breakpoints at upstream and downstream of the SRY gene region. We conclude that the testicular development in 46,XX male patients were due to insertion or translocation of SRY gene into X chromosome or autosomes.

• Journal of Animal Science and Technology
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• v.47 no.3
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• pp.317-324
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• 2005

A method for sex determination of pigs was examined using polymerase chain reaction(PCR). Sex determining region Y(SRY) gene encoded on Y chromosome plays a key role for primary male development. Zinc finger X-Y(ZFX-ZFY) gene, one of the X-V homology gene group was found on the X and Y chromosomes, respectively, We tested for molecular sexing by amplification patterns of SRY and ZF genes. Genomic DNAs from various resources including porcine hairs and semen collected from domestic pig breeds and native pigs was used for PCR assay of each gene. The amplified products for porcine SRY gene were yielded only in males but not in females. On the other hand, two differential patterns were observed in amplification of ZF gene reflecting the chromosomal dimorphism by a length polymorphism between X and Y chromosomes. Of both, a common band was detected in all individuals tested so that this band might be amplified from ZFX gene as a PCR template, but another is specific for males indicated that from ZFY. The result of PCR assay provides identical information to that from investigation of phenotypic genders of the pigs tested. We suggest that this PCR strategy to determine porcine sexes using comparison of the amplification patterns of the SRY gene specific for Y chromosome and the dimorphic ZF gene between X and Y chromosomes may be a rapid and precise method for discrimination of two sexes and applied to DNA analysis of small samples such as embryonic blastomere, semen, and hairs.