• Title/Summary/Keyword: Molecular Sex Determination

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Molecular Sex Determination Using Sexual Dimorphisms between ZFX and ZFY Genes in Korean Hares(Lepus coreanus Thomas) (한국멧토끼 ZFX와 ZFY 유전자의 성별 이형성과 분자 성판별)

  • Han, Sang-Hyun;Cho, In-Cheol;Lee, Sung-Soo;Oh, Moon-You;Oh, Hong-Shik
    • 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.

Typing of DNA Extracted from Cigarette Butts for Individual Identification (타액반 피검물에서 개인식별을 위한 DNA의 유전자형 검사)

  • Kyong-Kyue Yoon;Juck-Joon Hwang;Chong-Youl Kim
    • Journal of Oral Medicine and Pain
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    • v.19 no.2
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    • pp.205-219
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    • 1994
  • Cigarette butts from 5 smokers were gathered and then, placed in room temperature for 1, 3, 5, 7, 15 days. The possible use of the cigarette butts for individual identification was evaluated in sex determination, amplification of D1S80 locus, polymorphisms of HLA-DQA1 gene from the extracted DNA. 1. DNA extraction was possible in cigarette butts weree left in room temperature for 15days, so it can be applicatable to individual identification by polymerase chain reaction(PCR). 2. Amplification of X-Y homologous amelogenin gene by PCR made it possible to identify the sex in saliva stains (cigarette butts). 3. Amplification of D1S80 locus can be acquired from adding the boving serum albumin and hot start PCR procedures from forensic samples such as saliva stains (cigarette butts), so the AMP-FLPs examining is possible. 4. Genotype could be determined simply and rapidly using Amplitype$TM$ HLA-DQ$\alpha$ forensic kit in examining the HLA-DQA1 gene. From the investigation, DNA extraction, sex determination, amplification of D1S80 locus, polymorphisms of HLA-DQA1 gene was successfully done even though the cigarette butts were left for 15 days at room temperature. Therefore cigarette butts are highly reliable and applicatable as molecular biologic samples for individual identification.

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Identification and molecular characterization of doublesex and mab-3-related transcription factor(dmrt) in brackish water flea, Diaphanosoma celebensis, exposed to bisphenol analogs

  • Cho, Hayoung;Jeon, Min Jeong;Lee, Young-Mi
    • Korean Journal of Environmental Biology
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    • v.39 no.2
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    • pp.160-168
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    • 2021
  • Doublesex and mab-3 related transcription factor(dmrt) play crucial roles in sex determination and sex differentiation in vertebrates and invertebrates. Although dmrt genes have been identified in vertebrates, little is known about aquatic invertebrates. In this study, two dmrt genes, namely, Dc_dmrt93B and Dc_dmrt99B, were identified from brackish water flea, Diaphanosoma celebensis. Transcriptional changes were observed in the dmrt genes when the flea was exposed to bisphenol(BP), an endocrine disruptor. Sequence and phylogenetic analyses showed that both dmrt genes contained two conserved domains, namely, DM and DMA, closely clustered with those of Daphnia spp. Additionally, a significant increase in the Dc_dmrt99B mRNA expression level was observed upon exposure to intermediate concentrations of BP (bisphenol A>bisphenol S=bisphenol F, p<0.05), while the expression of Dc_dmrt93B mRNA was slightly modulated. These findings imply that the two dmrt genes may be involved in sex differentiation of D. celebensis. Furthermore, it was found that the ability of BP to modulate dmrt genes could affect development and reproduction. This study provides a basis for understanding the function of the dmrt genes and the molecular mode of action of BP in small crustaceans.

Molecular Sexing Using SRY and ZF Genes in Pigs (돼지 SRY와 ZF 유전자를 이용한 성판별 기법)

  • Cho, I.C.;Kang, S.Y.;Lee, S.S.;Choi, Y.L.;Ko, M.S.;Oh, M.Y.;Han, Sang-Hyun
    • 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.

Identification of RAPD markers linked to sex determination in guggal [Commiphora wightii (Arnott.)] Bhandari

  • Samantaray, Sanghamitra;Geetha, K.A.;Hidayath, K.P.;Maiti, Satyabrata
    • Plant Biotechnology Reports
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    • v.4 no.1
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    • pp.95-99
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    • 2010
  • Decamer RAPD primers were tested on dioeceious and hermaphrodite plants of Commiphora wightii to identify sex-specific molecular markers. Sixty different random decamer primers were screened out of which only three primers were found to be associated with sex expression. A ~1,280-bp fragment from the primer OPN06 was found to be present in all the female individuals. Another primer OPN 16 produced a unique ~400-bp amplification product in only hermaphrodite individuals. The third marker, OPA20 amplified a ~1,140-bp fragment from female and hermaphrodite DNAs, but failed to do so from the male plant DNAs.

Identification of Female Specific Genes in the W Chromosome that are Expressed during Gonadal Differentiation in the Chicken

  • Rallabandi, Harikrishna Reddy;Yang, Hyeon;Jo, Yong Jin;Lee, Hwi Cheul;Byun, Sung June;Lee, Bo Ram
    • Korean Journal of Poultry Science
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    • v.46 no.4
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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.

The Sex Determination Mechanisms in Maize: Cell Death, Cell Protection and Cell Cycle Arrest (옥수수 성 결정 메커니즘: 세포 사멸, 세포 방어, 세포주기 멈춤)

  • Kim, Jong-Cheol;Lee, Kyun-Oh
    • Journal of Life Science
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    • v.16 no.4
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    • pp.699-703
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    • 2006
  • Maize (Zea mays L.) is a monoecious plant, which separates male (tassel) and female (ear) floret that evolved into increasing heterogeneity. In each floret, male or female, bears both one pistil and three stamens primodia before diverged to unisexual state. When diverged to tassel, pistil cell death occurs in the pistil primodium, which is mediated by TASSELSEED genes. In contrast, cell protection occurs in the ear pistil from TASSELSEED-mediated cell death, which is mediated by SILKLESS1 gene. On the other hand, cell cycle arrest occurred for a long time in the ear stamens and then the stamens eventually dye. The cell cycle regulating genes such as CYCLIN B and WEE1 are involved in this process. Furthermore, the temporal and spatial regulation of gibberellin biosynthesis may cause cell cycle block in arresting stamen cells. This review describes the cell death, cell protection, and cell cycle arrest mechanism during maize sex determination process at the molecular, cellular and developmental biology, and genetic levels.

Identification of Sex-specific Expression Markers in the Giant Tiger Shrimp (Penaeus monodon)

  • Khamnamtong, Bavornlak;Thumrungtanakit, Supaporn;Klinbunga, Sirawut;Aoki, Takashi;Hirono, Ikuo;Menasveta, Piamsak
    • BMB Reports
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    • v.39 no.1
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    • pp.37-45
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    • 2006
  • Bulked segregant analysis (BSA) and AFLP were used for isolation of genomic sex determination markers in Penaeus monodon. A total of 256 primer combinations were tested against 6-10 bulked genomic DNA of P. monodon. Five and one candidate female- and male-specific AFLP fragments were identified. Female-specific fragments were cloned and further characterized. SCAR markers derived from FE10M9520, FE10M10725.1, FE10M10725.2 and FE14M16340 provided the positive amplification product in both male and female P. monodon. Further analysis of these markers using SSCP and genome walk analysis indicated that they were not sex-linked. In addition, sex-specific (or differential) expression markers in ovaries and testes of P. monodon were analyzed by RAP-PCR (150 primer combinations). Twenty-one and fourteen RAP-PCR fragments specifically/differentially expressed in ovaries and testes of P. monodon were successfully cloned and sequenced. Expression patterns of 25 transcripts were tested against the first stranded cDNA of ovaries and testes of 3-month-old and broodstock-sized P. monodon (N = 5 and N = 7 - 10 for females and N = 4 and N = 5 - 7 for males, respectively). Five (FI-4, FI-44, FIII-4, FIII-39 and FIII-58) and two (M457-A01 and MII-51) derived RAP-PCR markers revealed female- and male-specific expression patterns in P. monodon. Surprisingly, MII-5 originally found in testes showed a higher expression level in ovaries than did testes of juvenile shrimps but a temporal female-specific pattern in P. monodon adults.

Personal identification of the excavated ancient human bone through molecular-biological methods (분자생물학적 방법을 통한 출토인골의 개인 동정-사천 늑도 출토 인골과 민통선 민묘 출토 인골을 중심으로)

  • Seo, Min-Seok;Lee, Kyu-Shik;Chung, Yong-Jae;Lee, Myeong-Hui
    • 보존과학연구
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    • s.22
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    • pp.27-40
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    • 2001
  • DNA typing is often used to determine identity from human remains. Recently, the molecular biological analysis of ancient deposits has become possible since methods for the recovery of DNA conserved in bones or teeth from archaeological remains have been developed. In the field of archaeology, one of the most promising approaches is to identify the individuals present in a mass burial site. We performed nuclear DNA typing and mitochondrial DNA sequencing analysis based on PCR from a Korea ancient human remain excavated from Sa-chon Nuk-island and civilian access controlline(CACL). A femur bone were collected and successfully subjected to DNA extraction, quantification, PCR amplification, and subsequently typed for several shot tandem repeat(STR)loci. 4 types of STR systems used in this study were CTT multiplex(CSF1PO, TPOX, TH01), FFv multiplex(F13A01, FESFPS, vWA), Silver STRⅢ multiplex(D16S539, D7S820, D13S317), and amelogenin for sex determination. This studies are primarily concerned with the extraction, amplification, and DNA typing of ancient human bone DNA samples. Also, it is suggestive of importance about closely relationship between both fields of archaeology and molecular biology.

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