• Clinical and Experimental Reproductive Medicine
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• v.21 no.3
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• pp.281-284
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• 1994

Sex determination in genomic DNA from human blood leucocytes was performed by amplification of human Y chromosome-specific DNA sequences using PCR technique. A clear DNA fragment(154 nucleotides long) was appeared only in the male genomic DNA, but no specific band was observed in the case of female genomic DNA and negative control. To know the sensitivity of this method, the amplification reaction was performed in genomic DNA diluted to 2pg equivalent to the amonut present in the single human cell, and clear band also observed. The PCR amplification was so succesfully performed in the single leucocyte separated from human blood using micromanipulator that this techniqe is assumed to be applied to single blstomere before embryo transfer.

• Korean Journal of Animal Reproduction
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• v.20 no.4
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• pp.443-451
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• 1997

These studies were conducted to determine the sex of preimplantation Hanwoo embryos produced in vitro using polymerase chain reaction(PCR). Y chromosome specific and bovine speicific DNA primers were synthesized and tested for embryo sexing. Bovine IVF embryos were produced in TCM 199 and CR1aa medium, and classified by developmental stages on Day 7 to 9. The effects of developmental rates to bovine IVF blastocysts on sex ratio were also investigated using PCR methods. The results obtained in this study were as follows; 1. Developmental rates to blastocyst from IVM/IVF embryos in TCM 199 and CR1aa medium for 9 days were 23.5 and 30.2%, respectively, and there was significant difference between the media(P<0.05). 2. Male to female ratio of early, mid, expanded and hatching balstocyst produced on Day 7 were 0.7:1, 1.4:1, 2.2:1, and 2.5:1, respectively, and male embryos was significantly higher proportion in expanding and hatching blastocysts(P<0.01). 3. On Day 8, male to female ratio of early, mid, expanded and hatching blastocysts were 0.6:1, 1:1, 2.5:1, and 2.7:1, respectively. Both expanded and hatching blastocysts obtained a significantly higher proportion of males(P<0.01). 4. The male : female ratio of early, mid, expanded and hatching blastocyst produced on Day 9 was 0.6:1, 0.8:1, 1:1, and 2.2:1, respectively. Hatching blastocysts had a significantly higher ratio of males(P<0.01). The developmental rate of IVM/IVF embryos to blastocyst for 9 day culture was higher in CR1aa than that in TCM 199 medium. For the sex ratio by developmental stages of IVF embryos, male ratio was higher in expanded blastocyst but female in early blastocysts.

• Journal of Life Science
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• v.20 no.12
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• pp.1902-1905
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• 2010

This study was conducted to increase the efficiency of farming practice in rainbow trout, Oncorhynchus mykiss, by sex reversal and chromosome-set manipulation techniques. Induction of sex-reversed gynogenetic diploid rainbow trout males and mass production of all-female rainbow trout by genetic sex reversal was performed. Phenotypic males in the gynogenetic diploid group were induced successfully by dietary administration of 5 mg of 17 alpha-methyltestosterone per kg diet for 82 days. All females were produced by crossing between normal female and sex-reversed gynogenetic diploid male rainbow trout.

• Asian-Australasian Journal of Animal Sciences
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• v.5 no.1
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• pp.45-49
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• 1992

Bovine drumstick of neutrophil leucocytes was studied on the quantitative and morphological characteristics and was evaluated as a diagnostic measure for bovine freemartin in newborn calves. Nuclear area of neutrophil (A, ${\mu}m^2$) and drumstick area (B, ${\mu}m^2$) were significantly correlated with average diameter of drumstick (ADD, ${\mu}m$) and following regression equations were obtained : $A=45({\pm}3)$ ADD-8, r = 0.74, $s.e.{\pm}0.6$, p < 0.01, $B=1.72({\pm}0.05)$ ADD-0.98, r = 0.93, $s.e.{\pm}0.1$, p < 0.01 Eight female siblings of heterosexual multiplets were diagnosed as freemartin from the results of chromosome analysis. Heterosexual multiplets had a very low frequency of drumstick in the nucleus of neutrophils irrespective of genetic sex. Diameters of drumstick fund in freemartin and male cotwin did not differ from those of normal cows. Examinations of drumstick in 800 neutrophils for both female and male siblings are concluded to be the best way to aid the detection of freemartinism of heterosexual twins at early life.

• Journal of Genetic Medicine
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• v.12 no.2
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• pp.118-122
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• 2015

Noninvasive prenatal test (NIPT) is a novel screening method for the diagnosis of fetal chromosomal aneuploidies. NIPT is based on technology that detects cell-free fetal DNA in maternal plasma and analyzes it with massively parallel sequencing technology to determine whether the fetus is at risk of trisomy 21, trisomy 18, trisomy 13 or sex chromosome abnormalities (SCAs). NIPT has been reported to have sensitivity of 99% and a false positive rate of less than 1% for detecting trisomy 21 and trisomy 18. Although extension of the application of NIPT to other SCAs has been attempted, there are concerns in extending NIPT to SCAs because of maternal or fetal mosaicism, undetected maternal SCAs, and multiple pregnancies. Recently, we assessed a pregnancy with the rare Turner syndrome mosaicism 45, X/47, XXX, which was reported as 45, X with NIPT. We present the case here and briefly review the current literatures on NIPT in testing for fetal monosomy X. To the best of our knowledge, this is the first report of the 45, X/47, XXX mosaicism in Korea to be reported as 45, X by NIPT with whole genome sequencing. This case report will provide valuable information for counseling women who want to undergo NIPT.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.9
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• pp.1227-1231
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• 2002

A powerful tool for chicken transgenesis could be established by employing a germline chimera production through primordial germ cell transplantation. This study was conducted to examine whether foreign gene-transfected gonadal primordial germ cells (gPGCs) have a migration activity into the gonad after transfer to recipient embryos. In Experiment 1, gPGCs of Korean Ogol Chicken were retrieved from 5.5-day-old embryos and subsequently transferred to the dorsal aorta of 2.5-day-old White Leghorn embryos after being labeled with PKH26 fluorescent dye. To confirm migration activity after transplantation, recipient embryos were sacrificed and examined on 3 days after transfer. Sex determination was concomitantly undertaken to examine whether sex of recipient embryos could affect the migration activity of gPGCs. All of embryonic gonads examined showed positive signals with PKH26 fluorescence and W-chromosome specific band by polymerase chain reaction (PCR) was detected in male embryos when gPGCs with ZW chromosome were transferred to recipient embryos. In Experiment 2, retrieved gPGCs were transfected with LacZ gene-containing cytomegalovirus promoter ($pCMV{\beta}$) by electroporation and subsequently transferred to recipient embryos. LacZ gene expression was identified in the gonads of 6 or 10-day-old recipient embryos and hatched-chicks. A total of 20 embryos and 12 hatched-chicks were examined and 11 of them (10 embryos and one hatched chicken; 11/32=34.4%) expressed $\beta$-galactosidase, a marker substance of LacZ gene. The results of this study demonstrated that foreign gene-transfected gPGCs can migrate and settle down into the gonad after being transferred into the blood vessel of the recipient embryos. This established technique will contribute to developing a peer biotechnology for transgenic chicken.

• Journal of Life Science
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• v.9 no.1
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• pp.69-75
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• 1999

Four species belonging to the Drosophila melanogaster complex were examined genetically and morphologically to analyze interspecific relationships. Insemination rates ranged from 96% to 99% within species crosses, but interspecific crosses among the four species exhibited a great variations in the frequency of successful matings. D. melanogaster females mated relatively well with males of other species and D. sechellia males were more successful in mating with females of other species. In the crosses among D. simulans, D. mauritiana and D. sechellia, hybrid flies were fertile in females, but sterile in males regardless of reciprocal matings. The phenogenetically relationship between this complex and their hybrids were investigated by the comparison of sex comb tooth number and genital arch of male. They were controlled by polygenic factors on the chromosome of both parents. The effects of temperature on viability of hybrids between D. melanogaster females and D. simulans males were investigated for detection of genes concerning the speciation. The temperature sensitivity of the hybrid was mainly controlled by genes located on the X chromosome of D. simulans males.

• Animal cells and systems
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• v.5 no.3
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• pp.237-241
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• 2001

Three STR loci (STRX1[AGAT]$_{n}$, HPRTB[AGAT]$_{n}$ and ARA[AGC]$_{n}$) on X chromosome and three other STR loci (DYS390[CTG(A)T]$_{n}$, DYS392[ATT]$_{n}$ and DYS39［GATA]$_{n}$) on Y chromosome were analyzed in 154 unrelated healthy Korean subjects. Four loci (STRX1, HPRTB, DYS390 and DYS393) were amplified by quadruplex polymerase chain reaction (PCR) using fluorescent labeled primers (FLP). ARA and DYS392 were amplified separately using single PCR, similarly by using FLP. They were then run in an automated DNA sequencer and were analyzed with Genescan software. We found 7 alleles (308-332 bp) in STRX1, 7 alleles (275-299 bp) in HPRTB, 16 alleles (252-315 bp) in ARA, 6 alleles (203-223 bp) in DYS390, 7 alleles (245-263bp) in DYS392 and 5 alleles (116-132 bp) in DYS393. The ＊13(34%), ＊13(5l%), ＊23 (l8%), ＊23(50%), ＊14(39%) and ＊13(40%) alleles were observed to be the highest frequencies of STRX1, HPRTB, ARA, DYS390, DYS392 and DYS393, respectively. The detection of STR loci on sex chromosomes by quadruplex PCR allows simple determination of sex and identification of an individual. individual.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.8
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• pp.1107-1112
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• 2009

The separation of X and Y chromosome-bearing sperm is of use in many aspects of livestock maintenance. In this study, we sought to determine the difference in DNA content between X- and Y-bearing sperm, separate sperm into X- and Y-enriched pools, and assess the efficacy of sorting. Sperm collected from Duroc and miniature pigs were stained with 20.8 $\mu{M}$ Hoechst 33342 and analyzed using a high-speed cell sorter. Measurement of the fluorescence intensity of stained sperm nuclei revealed that the X-bearing sperm of Duroc and miniature pigs respectively contain 2.75% and 2.88% more DNA than Y-bearing sperm. In total, 50.18% of the sperm were assigned to the X-sorted sample and 49.82% was assigned to the Y-sorted sample for Duroc pigs. For miniature pigs, the Xsorted sample represented 50.19% of the population and the Y-sorted represented 49.81% of the population. Duplex PCR was used to evaluate accuracy of sorting. A fast and reliable method for porcine sexing was developed through amplification of the sex-determining region of the Y chromosome gene (SRY). Oligonucleotide primers were designed to amplify the conserved porcine SRY high motility group (HMG) box sequence motif. We found that the primer pair designed in this study was 1.46 times more specific than previously reported primers. Thus, this study shows that the present method can be applied in porcine breeding programs to facilitate manipulation of the sex ratio of offspring and to achieve precise sexing of porcine offspring by amplification of the HMG box of the SRY gene.

• Journal of Aquaculture
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• v.9 no.1
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• pp.101-106
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• 1996

Gynogenetic males induced from sex reversed female (${\Delta}XY$) were crossed with normal female (XX) for analysing their genotypes. The fish tested produced a high percentage of male progenies (93.3 to $100\%$) and were considered as supermales (YY). Superfemales (${\Delta}YY$) were also produced by combination of sex reversal and chromosome manipulation techniques. Superfemale fish can be produced approximetly $90\%$ of male when the fish were crossed with normal male. Chi-square values against an expected 1 : 1 (male : female) ratio were highly significant for both YY males${\times}$ normal females (P<0.01 or P<0.001) and ${\Delta}YY$ females${\times}$normal males (P<0.005 or P<0.001). All male progenies were produced consistently when crossed supermales (YY) with superfemales (${\Delta}YY$).