• Journal of Embryo Transfer
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• v.24 no.3
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• pp.225-230
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• 2009

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.

• Clinical and Experimental Pediatrics
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• v.48 no.12
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• pp.1317-1323
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• 2005

Purpose : This study was performed to evaluate the recent frequency of karyotypes in different sex chromosome abnormalities and to evaluate the age and clinical manifestations at diagnosis. Methods : Peripheral blood leukocytes were obtained from subjects who were clinically suspected to have sex chromosome abnormalities and referred to the cytogenetic laboratory in the Department of Pediatrics, Kyungpook National University Hospital from February 1981 to August 2001. Results : The relative frequencies of different sex chromosome abnormalities were Klinefelter(52 percent), Turner(42 percent), XXX syndrome(3 percent) and mixed gonadal dysgenesis(3 percent). The populations of different karyotypes in Klinefelter syndrome were 47,XXY(97 percent) and 46,XY/47,XYY(3 percent). The populations of different karyotypes in Turner syndrome were 45,X(67 percent,), mosaicism(23 percent), and structural aberrations(10 percent). The populations of different karyotypes in XXX syndrome were 47,XXX(67 percent,) and 46,XX/47,XXX(33 percent). All mixed gonadal dysgenesis were 45,X/46,XY. Eighty one percent of sex chromosome abnormalities was diagnosed after puberty. Patients diagnosed with Klinefelter and Turner syndrome in infancy showed nearly normal phenotypes or had minor congenital malformations. Conclusion : Early diagnoses of sex chromosome abnormalities is required to prevent associated morbidities and to maximize growth and development. We have to pay careful attention in diagnoses of Turner syndrome because of the high proportion of mosaicism and structural aberrations.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.11
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• pp.1689-1693
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• 2007

The objective of this study was to develop a simplified, efficient, and accurate protocol for sexing goat embryos. Based on the amelogenin gene located on the conservation region of X- and Y- chromosomes, a pair of primers was utilized and the system of PCR was established to amplify a 262 bp fragment from the X- chromosome in female goats, and a 262 bp fragment from X- chromosome and 202 bp fragment from the Y- chromosome in male goats, respectively. The accuracy and specificity of the primers were assessed using DNA template extracted from goat whole blood sample of known sex. 100% (10/10) concordance was obtained by using the PCR assay. Fifty-one biopsied embryos were transferred into 25 recipient goats on the same day that the embryos were collected and sex of the kid was confirmed after parturition. Eighteen kids of predicted sex were born. The biopsied samples from 51 goat embryos were amplified with 100% efficiency and 94.7% accuracy. In conclusion, our results indicated that PCR sexing protocols based on the amelogenin gene is highly reliable and suitable for sex determination of goats.

• The Korean Journal of Zoology
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• v.39 no.1
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• pp.75-88
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• 1996

OTF9-63 (OTF9) and P19S1O1A1 (P19) embryonal carcinoma (EC) cells were examined for their ability to produce the readivation of inactive X chromosomes from somatic cells. They were hybridized with various somatic cells and resulting HATr EC-somatic cell clones were analysed for their morphology, chromosomal replication pafterns and expression proffies of X-linked and distantiy located genes, Hprt and Pgk-1. The results demonstrated that 0RF9 cells could reactivate the inactive X chromosome whereas P19 cells could not. In adition, EC-somatic cell hybrids tended to reduce the number of sex chromosomes in long-term culture, resulting m 1:2 ratio of sex chromosomes to autosomes The use of EC cell hybrids provides an experimental system for studying the mechanism(s) of the X-reactivatio that is initiated and maintained from meiotic prophase of oogenesis to early embryogenesis.

• Korean journal of applied entomology
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• v.35 no.3
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• pp.216-220
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• 1996

Drosophila simulans and D. mauritiana are sibling species, the former cosmopolitan and the latter restricted to the oceanic island of Mauritius. Sex comb-tooth number of male flies of D. simulans were about 9.83, while those of D. mauritiana were 12.90. Genital arch of D. simulans is large semicircular shaped expasion, while that of D. mauritiana is a narrow fingerlike expansion. We used classical genetic analysis to measure effects of genes on the X chromosome responsible for numeral and morphological differences in sex comb-tooth and genital arch between these species, respectively. For these purposes, mutant strain of D. simulans and wild type strain of D. mauritiana were hybridized and males of the FI and the backcrossed progenies were compared with two characters above mentioned. The sex comb-tooth number of F, males were about 11.79, and the genitalia of F, male were intermediate in shape between those of D. simulans and D. mauritiana. Genetic analysis of sex comb-tooth number and genital arches differing between D. simulans and D. mauritiana showed that very little diffemce was due to effect of the X chromosome.

• International Journal of Industrial Entomology and Biomaterials
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• v.8 no.2
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• pp.161-167
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• 2004

Silkworms sex determination drew high attention from researchers. Sex chromosomes on the silkworm are of ZW type for females and ZZ type for males. Chromosome W plays an important role in sex determination. Although several molecular linkage maps have been constructed for silkworm, very few markers are discovered on the W chromosome. In order to look for molecular markers and to further locate the Fern gene on chromosome W, we used genomic DNA from both female and male larvae of a silkworm strain named 937 as PCR templates for RAPD amplification with 200 arbitrary 10-mer primers. The amplification results showed three female-specific bands, namely ${OPG-07_496}, {OPC-15_1,660} and {OPE-18_1,279}$. Further verification, however, revealed no band from OPG-07 and OPC-15 in either sex in the strain 798, but OPE-18 provided female-specific band in the strains Suluan7 and C108, and absent in both males and strain 798. This indicates that the bands from ${OPG-07_496} and {OPC-15_1,660}$ are probably female-specific in strain 937, and the band from OPE-18 was probably amplified from a common segment shared by most strains. The genomic DNAs from OPG-07 and OPC-15 were cloned and sequenced. Sequence analysis showed that the DNAs from OPG-07 and OPC-15 have high identities with the retrotransposable elements, and DNA from OPC-15 contains a portion of sequence which probably encodes an eukaryotic translation initiation factor 4E binding protein (eIF4EBP).

• Korean Journal of Animal Reproduction
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• v.20 no.2
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• pp.179-190
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• 1996

Sexing and developing from splitted embryos which were fertilized in vitro implicate a possibility of production of the superior and sex controlled individuals. This study was carried out to investigate the production of transferable late blastocysts from in vitro fertilized embryos and to analyze sex by chromosome analysis from same embryos. In results, the ratio of cleavage and fertility of bovine follicular oocytes matured in vitro was 90% in co-cultured with granulosa cells. The competence of embryonic development from in vitro matured and fertilized bovine oocytes was 38% in co-cultured with bovine oviductal epithelial cells. To produce a lot of transferable embryos, therefore, the best conditon of culture system was co-cultured with granulosa cells for immature bovine oocytes and then co-cultured with bovine oviductal eptithelial cells for matured and fertilized oocytes. In chromosome analysis, 93% of in vitro fertilized embryos were very important aspect in chromosome preparation from bovine embryos such as duration of colcemid treatment, weakening of zona pellucida, methods of hypotonic treatment and fixation treatment.

• 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.

• The Korean Journal of Zoology
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• v.7 no.1
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• pp.29-32
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• 1964

A study on chromosome of leucocytes in blood cultures derived from 6 normal Korean was performed . Exact chromosome counts were carried out on 205 cells in male, 211 in female , of which 86.05% revealed a chormosome mordal number of 46. On the basis of relative chromosome lengths and position of centromeres, the Karyotype that the human chromosomes were classified into 7 groups with 22 airs of autosome and one pair of sex chromosome was determined accoridng to the method of denver report. The chromosome number on metaphase was observed in short term cultures of leucocytes from the peripheral blood of 2 patients with chronic granulocytic leukemia and 1 patient with acute granulocitic leukemia . and the chromosome morpholoogy was also investigated in one acute leukemic patient. In all leukemic cases the leucocytes showed the constant value of 46 in the stem -line of chormosome number. But the frequency of cells with 46 chromosomes appeared in the 3 cases was 67.30% in average with a slightly higher range in hypo-andhyper-diploid chromosome numbers than in normal human, The idiogram analysis did not show any abnormality of chromosome in acute leukemic cells.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.11
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• pp.1531-1535
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• 2002

It has been known that the sex of chicken cells can be most accurately identified by fluorescence in situ hybridization (FISH). However, the presently available FISH has not been widely used for sex identification, because the procedures for cell preparation and FISH itself are complicated and time-consuming. The present study was undertaken to test a rapid FISH procedure for sexing chicken. A FISH probe was simultaneously synthesized and labeled with digoxigenin by polymerase chain reaction (PCR) targeting a 416 bp segment of the 717 bp XhoI family fragment which is repeated over 10 thousand times exclusively in the W chromosome. Sexing by FISH was performed on cytological preparations of early embryos, adult lymphocytes and feather pulps of newly hatched chicks. The DNA probe hybridized to all types of uncultured interphase as well as metaphase female but not male cells that had been examined. Moreover, consistent with the known site of the XhoI family, the hybridization signal was localized to the pericentromeric region of the W chromosome. We, therefore, conclude that the present PCR-based FISH can be used as a rapid and reliable sex identification procedure for chicken.