• Clinical and Experimental Reproductive Medicine
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• 제21권3호
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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.

• 한국가축번식학회지
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• 제20권4호
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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.

• 생명과학회지
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• 제20권12호
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• pp.1902-1905
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• 2010

본 연구는 무지개송어 양식 산업의 생산성 향상을 위한 일환으로 염색체공학 기법을 이용하여 전 암컷 무지개 송어의 대량생산을 유도 하였다. 자성발생 2배체를 유도한 후 17 alpha-methyltestosteron으로 성전환을 성공적으로 유도하였다. 성전환된 수컷에서는 일반 암컷 모양을 띤 생식소가 형성 되었으나, 수정관의 발달은 보이지 않는 전형적인 성전환 개체의 특징을 나타내었다. 성전환된 자성발생 2배체 가짜수컷을 이용하여 정상 암컷과 단순교배로 발안율 55.7%, 부화율 52.9%의 전 암컷 집단 97,850 마리를 얻을 수 있었고, 4개월 사육 후 치어기(6~7 cm)때의 생식소 확인 결과 100% 암컷임이 확인되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제5권1호
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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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• 제12권2호
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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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• 제15권9호
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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.

• 생명과학회지
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• 제9권1호
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• pp.69-75
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• 1999

Genus Drosophila에서 melanogaster complex내의 4종에 대한 유연관계 해석을 위하여, insemination test에 의한 premating isolation, 잡종형성 여부에 의한 postmating isolation, 형태 형성에 관한 영향 및 종 분화 관련 유전자의 특징 등을 대상으로 조사하였다. 종내 교배에서는 insemination rate는 96∼99% 정도였고 종간 교배에서는 정역 교배간에 심한 변이를 보였으며 D. melanogaster 암컷은 타종 수컷과의 교배에서 전반적 성공률이 높으며 D. sechellia 수컷은 타종 암컷과의 교배에서 비교적 높은 교배 성공률을 보였다. 종간 잡종 형성에서는 특히 simulans, mauritiana 및 sechellia 사이에서 임성이 완전한 암컷과 불임의 수컷이 형성되어 이들 3종이 더욱 근연임을 시사하고 있다. sex comb과 genital arch에 대한 잡종의 형태 형성에 관한 영향은 대부분이 polygene에 의하여 조절되고 있음을 알 수 있었다. D. melanogaster와 simulans에 있어서 분화 관련 유전자로 추정되는 잡종의 온도 감수적 생존도는 주로 simu-lans의 X 염색체상의 유전자에 의하여 조절되고 있음이 분석되었다.

• Animal cells and systems
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• 제5권3호
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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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• 제22권8호
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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.

• 한국양식학회지
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• 제9권1호
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• pp.101-106
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• 1996

XY 암컷에서 유도된 자성발생성 2배체 수컷 및 이의 성전환 암컷으로 부터 YY수컷(초수컷) 및 ${\Delta}YY$ 암컷(초암컷)을 선발하기 위하여 각각 정상 암컷 및 정상 수컷과 교배하여 자손 검정을 실시하였다. 교배가 이루어진 자성발생성 이배체 수컷 7마리 중 3마리가 YY 수컷으로 판정되었으며, 이들 자손의 수컷율은 $93.3\%\~100.0\%$로서 정상 교배에서 예상되는 성비인 1 : 1에 대한 $X^2$ test 결과 매우 유의하게 나타났다(P<0.01 또는 P<0.001). 자성발생성 2배체 성전환 암컷의 경우 교배가 이루어진 3마리 중 2마리가 ${\Delta}YY$ 암컷으로 판정되었으며, 이들 자손의 수컷율은 각각 $92.9\%$ 및 $86.2\%$ 그리고 $87.5\%$ 및 $95.0\%$로서 1 : 1 성비에 대해 매우 유의하였다(P<0.005). YY 수컷과 ${\Delta}YY$ 암컷간의 교배에서 전수컷 자손만이 생산되었다.