• 한국축산식품학회지
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• 제27권3호
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• pp.351-356
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• 2007

본 연구는 포유류의 Y-염색체상에 존재하는 SRY 및 ZFY의 웅성 특이적 성 결정 유전자를 이용하는 PCR 기법으로 쇠고기 성판별 기술을 개발하고자 수행하였다. 성 결정 유전자 영역의 특정 염기서열을 포함하는 primer를 각각 설계 합성하고 이들 primer를 이용하여 PCR증폭을 실시한 다음, 각각의 증폭산물을 1.5% agarose gel에 전기영동하여 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 숫소에서 유래한 쇠고기에서 웅성 특이적인 DNA band가 정확히 검출된 반면 암소에서 유래한 쇠고기에서는 웅성 특이적 DNA band가 전혀 검출되지 않았다. 또한 쇠고기 성 감별법을 도축 후 가공 유통판매단계에 실제 활용 가능성을 검증하고자 시중에 유통되고 있는 등심 및 갈비 포장육 350시료를 무작위 추출하여 성 판별을 실시한 결과 숫소가 252시료(72%) 그리고 암소가 98두(28%)로 조사되었다. 따라서 본 연구에서 개발한 SRY 또는 ZFY의 웅성 특이적 성 결정 유전자를 이용하는 쇠고기 성 감별기술은 생산단계는 물론 도축 후 가공 유통 판매되고 있는 모든 쇠고기의 암수 성감별을 위한 유용한 DNA 표지인자로 활용할 수 있을 것으로 기대된다.

• Journal of Animal Science and Technology
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• 제54권3호
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• pp.219-226
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• 2012

Gender specificity in muscle growth and development is well known. Genesis of muscle is dependent on proliferation and differentiation potential of resident myogenic satellite cells (MSCs) present in muscle fibers. Multipotential capacity of forming myocyte, osteocyte, and adipocyte like cell makes MSCs a unique stem cell. To understand the molecular mechanism involved in determination of muscle quality due to difference in hormone concentration of different gender of animals, MSCs were isolated from bovine skeletal muscle and cultured in male, female, and castrated serum supplemented media. DNA microarray used consisted of 24,000 spots with 70 mer oligo in each spot. A total of 88 genes were up-regulated and 551 genes were down-regulated by more than two fold. Among up-regulated gene, 33, 34, and 21 genes were found up-regulated in cells grown in male, female, and castrated serum, respectively. Interestingly, male serum showed 4, female 11 and castrated male showed 4 genes expressed highly in each gender. Further study on the highly up-regulated gene may unfold the mystery of gender specificity found in muscle development. Also, the identification of differentially expressed genes in gender-specific serum will add information on infrastructure of bovine genome research.

• 한국동물생명공학회지
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• 제34권2호
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• pp.111-116
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• 2019

This study was conducted to analyze the specific genes associated with sex-determination in Korean native cow. The highly organized spermatogenesis requires accurate spatial and temporal regulation of gene expression, which is governed by transcriptional, post-transcriptional, and epigenetic processes. Recently, farmers have been interested in determining the sexual identity of the calves in their farm. We analyzed the sperm of Korean native and Holstein cows, which were supplied from Hanwoo Improvement Center. We evaluated sperm motility and expression of sperm-specific genes after treating semen with both male- and female reagents. Sperm motility in Korean native cows decreased by approximately 10% in the first 30 minutes after treatment with sex-determination reagent. However, sperm motility of Holstein cows decreased to 60-70% after 15 minutes and to 20-30% after 30 minutes. We selected six specific genes expressing in the spermatozoa to analysis the gene expression level. The Real-time PCR results suggest that the selected genes (Gimap4, Tmeff1, Rac2, Abi2, Rac1, and Clu) were highly expressed in the group treated with the male reagent compared to the group treated the female reagent and to the untreated-group (control). In the present study, we suggest that the selected genes play a pivotal role in sex-determination.

• 한국가축번식학회지
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• 제20권2호
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• pp.89-99
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• 1996

The purpose of this study was to develop the diagnosis techniques for sex determination of rabbit embryos at preimplantation stage. To detect male specific sequences using polymerase chain reaction, two genes functional on sex determination including SRY and ZFX/Y genes were targeted using multiple oligonucleotide primer sets. Three of them for conserved SRY gene were used for appropriate amplification pattern, and then only one primer set #3 proved to be most efficient, showing male-specific strong signal ofamplified sequences. Using this male specific bandsfrom human, cattle, pig and mouse, the gender of rabbit was determined. As an another system for sex determination system, amplified 910bp fragment from ZFX/Y was digested with several restriction endonuclease and showed gender specific restriction fragments only by Hinf I. Using two different system for sex identification of rabbit in this study, blind tests for 17 samples was conducted and showed identical results from two different methods. And then, amplification limit of PCR reaction for template DNA was estimated using various amounts of DNA for both SRY and ZFX/Y systems, resulted as 20pg and 800pg, respectively. With this results, test for gender identification of rabbit embryos were performed using SRY derived amplification system. From total 22 embryos selected for its developmental state 18 were identified as male embryos, showing significant difference from expected sex ratio 1:1. This biased sex ratio was interpreted as to have been caused by the fact, reported by the fact, reported by several researchers, that male embryos develop more rapidly and are more resistant against the in vitro manipulation than female embryos.

• 한국가금학회지
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• 제46권4호
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• pp.287-296
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• 2019

Avian sex determination system involves the male ZZ and female ZW chromosomes. However, very few studies are reported the expression, functional role and importance of genes on the W chromosome because of its small and highly heterochromatic genomic regions. Recent studies demonstrated that the W chromosome may have critical roles in physiology, sex determination and subsequent sexual differentiation in chickens. Therefore, gene annotation, including describing the expression and function of genes in the chicken W chromosome, is needed. In this study, we have searched the W chromosome of chickens and selected a total of 36 genes to evaluated their specific expression in the testis and ovary at various developmental stages such as embryonic day 6 (E6), hatch and adult. Interestingly, out of 36 genes in chicken W chromosome, we have found seven female-specific expression at E6.5 day, indicating that they are functionally related to female chicken gonadal differentiation. In addition, we have identified the stage specific gene expression from the sex specific genes. Furthermore, we analyzed the relative location of genes in the chicken W chromosome. Collectively, these results will contribute molecular insights into the sexual determination, differentiation and female development based on the W chromosome.

• Genomics & Informatics
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• 제2권1호
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• pp.30-35
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• 2004

To investigate the XIST gene expression and its effect in a Klinefelter's patient, we used Klinefelter's syndrome (XXY) patient with azoospermia and also used a normal male (XY) and a normal female (XX) as the control, We were performed cytogenetic analysis, Y chromosomal microdeletion assay (Yq), semi-quantitative RT-PCR, and the Northern blot for Klinefelter's syndrome (KS) patient, a female and a male control, We extracted total RNA from the KS patient, and from the normal cells of the female and male control subjects using the RNA prep kit (Qiagen), cDNA microarray contained 218 human X chromosome-specific genes was fabricated. Each total RNA was reverse transcribed to the first strand cDNA and was labeled with Cy-3 and Cy-5 fluorescein, The microarray was scanned by ScanArray 4000XL system. XIST transcripts were detected from the Klinefelters patient and the female by RT-PCR and Northern blot analysis, but not from the normal male, In the cDNA microarray experiment, we found 24 genes and 14 genes are highly expressed in KS more than the normal male and females, respectively. We concluded that highly expressed genes in KS may be a resulted of the abnormal X inactivation mechanism.

• Clinical Psychopharmacology and Neuroscience
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• 제20권3호
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• pp.474-481
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• 2022

Objective: In this study, we investigated sex- and region-specific effects of acute trimethyltin (TMT) exposure on mitochondrial biogenesis. Methods: We treated TMT to primary neuronal cultures and 4-week-old male and female mice. We measured the mitochondrial DNA copy numbers using the quantitative polymerase chain reaction method. We also measured mitochondrial biogenesis related genes (sirtuin-1, estrogen-related receptor alpha, cytochrome C oxidase subunit IV) by western blotting. Results: The mitochondrial DNA copy number increased in the primary hippocampal neuron; however, it decreased in the primary cortical neuron. The mitochondrial copy number increased in the hippocampus and decreased in the cortex in the TMT treated female mice, though the mitochondrial copy number increased in both cortex and hippocampus in the TMT treated male mice. TMT treatment increased sirtuin-1 expression in the male hippocampus but did not in the female brain. In the female brain, estrogen-related receptor alpha expression decreased in the cortex though there is no significant change in the male brain. The protein level of mitochondrial protein, cytochrome C oxidase subunit IV, increased in both cortex and hippocampus after TMT injection in male mice brain, but not in female mice brain. Conclusion: Our data suggest that acute TMT exposure induces distinct sex-specific metabolic characteristics in the brain before significant sexual maturation.

• Animal cells and systems
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• 제11권2호
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• pp.165-168
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• 2007

The nucleotide sequences of a male-specific marker sex determining region Y (SRY) gene and a mitochondrial cytochrome B (CYTB) gene were characterized and analyzed to establish a molecular method for identification of species and sex of Korean roe deer (Capreolus pygargus tianschanicus). Similarity search result of SRY sequences showed very similar result to those reported in Moose (Alces alces) and Reindeer (Rangifer tarandus), both of which had 95.9% similarity in identity. CYTB genes were very similar to those reported in Siberian roe deer (C. pygargus pygargus) which had 98.6% similarity and not to European roe deer (C. capreolus), suggesting that the DNA samples tested were of Siberian roe deer lineage. Polymerase chain reaction (PCR)-based sex typing successfully discriminated between carcasses of male and female roe deer. Males had SRY band on agarose gels and females did not. The result of this molecular sex typing provided similar information with that obtained by genital organ observation. Therefore, this molecular method using male specific marker SRY and mitochondrial CYTB genes would be very useful for identification of the species and sex of the carcass remains of roe deer.

• 생명과학회지
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• 제13권6호
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• pp.958-969
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• 2003

인간 Y 염색체는 엄격히 부계 유전되고 그 길이의 대부분은 남성 감수분열 동안 교차가 일어나지 않는다. 비록 이영역이 비 재조합 영역 Y (non-recombining region Y: NRY)로 불려왔지만, 풍부한 재조합의 발견으로 그것은 남성 특이 영역 (male-specific region Y: MSY)으로 재 명명(命名)되었다. MSY는 이질염색질 (heterochromatin) 서열과 세가지 분류의 진정염색질 (euchromatin) 서열 (X-전위영역, X-퇴화영역, 증폭영역)이 모자이크화 되어있다. X-전위영역의 서열은 X 염색체의 상동 좌위와 약 99% 동일성을 가진다. X-퇴화영역 서열은 고대 상 염색체가 현대의 X와 Y 염색체로 진화되면서 남아 있는 부분이다. 증폭영역의 8개의 회문구조는 인간 Y염색체의 남성 특이영역의 4분의 1을 차지한다. 이들은 많은 정소 특이 유전자를 포함하고, 회문구조서열 사이의 상동성은 약 99.97%이다. 이 회문구조의 양쪽 팔은 계속되는 유전자 교환에 의해 유지되며, 서로 협력하여 진화된다. 새로 태어나는 남성당 평균 약 600 염기당 하나가 Y-Y유전자 교환을 겪고, 정소 특이적 다중유전자군의 진화에 중요한 역할을 한다.

• Clinical and Experimental Reproductive Medicine
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• 제40권3호
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• pp.107-114
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• 2013

Homeobox genes play essential roles in embryonic development and reproduction. Recently, a large cluster of homeobox genes, reproductive homeobox genes on the X chromosome (Rhox) genes, was discovered as three gene clusters, ${\alpha}$, ${\beta}$, and ${\gamma}$ in mice. It was found that Rhox genes were selectively expressed in reproduction-associated tissues, such as those of the testes, epididymis, ovaries, and placenta. Hence, it was proposed that Rhox genes are important for regulating various reproductive features, especially gametogenesis in male as well as in female mammals. It was first determined that 12 Rhox genes are clustered into ${\alpha}$ (Rhox1-4), ${\beta}$ (Rhox5-9), and ${\gamma}$ (Rhox10-12) subclusters, and recently Rhox13 has also been found. At present, 33 Rhox genes have been identified in the mouse genome, 11 in the rat, and three in the human. Rhox genes are also responsible for embryonic development, with considerable amounts of Rhox expression in trophoblasts, placenta tissue, embryonic stem cells, and primordial germ cells. In this article we summarized the current understanding of Rhox family genes involved in reproduction and embryonic development and elucidated a previously unreported cell-specific expression in ovarian cells.