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      형광접합보인법 (FISH)을 이용한 소 체외수정란의 성 감별 = Sex Determination of In Vitro Fertilized Bovine Embryos by Fluorescence In Situ Hybridization Technique

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      https://www.riss.kr/link?id=A104700438

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      다국어 초록 (Multilingual Abstract)

      Sexing from bovine embryos which were fertilized in vitro implicate a possibility of the sex-controlled cattle production. This study was carried out to investigate the possibility of determining of embryo sex by fluorescence in situ hybridization (FISH) technique. FISH was achieved in in vitro fertilized bovine embryos using a bovine Y-specific DNA probe which constructed from the btDYZ-1 sequences. To evaluate Y-chromosome specificity of the FISH probe, metaphase spreads of whole embryos and lymphocytes were prepared and tested. A male-specific signal was detected on 100% of Y chromosome bearing metaphase specimens. Using the FISH technique with a bovine Y-specific probe, 232 whole embryos of 8 cell- to blastocyst-stage were analyzed. Observing the presence of the Y-probe signal on blastomeres, 102 embryos were predicted as male, and 130 embryos as female. The determining rate of embryo sex by FISH technique was about 93% regardless of embryonic stages. In conclusion, the FISH using a bovine Y-specific DNA probe is an accurate, reliable and quick method for determining the sex of bovine embryos.
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      Sexing from bovine embryos which were fertilized in vitro implicate a possibility of the sex-controlled cattle production. This study was carried out to investigate the possibility of determining of embryo sex by fluorescence in situ hybridization (FI...

      Sexing from bovine embryos which were fertilized in vitro implicate a possibility of the sex-controlled cattle production. This study was carried out to investigate the possibility of determining of embryo sex by fluorescence in situ hybridization (FISH) technique. FISH was achieved in in vitro fertilized bovine embryos using a bovine Y-specific DNA probe which constructed from the btDYZ-1 sequences. To evaluate Y-chromosome specificity of the FISH probe, metaphase spreads of whole embryos and lymphocytes were prepared and tested. A male-specific signal was detected on 100% of Y chromosome bearing metaphase specimens. Using the FISH technique with a bovine Y-specific probe, 232 whole embryos of 8 cell- to blastocyst-stage were analyzed. Observing the presence of the Y-probe signal on blastomeres, 102 embryos were predicted as male, and 130 embryos as female. The determining rate of embryo sex by FISH technique was about 93% regardless of embryonic stages. In conclusion, the FISH using a bovine Y-specific DNA probe is an accurate, reliable and quick method for determining the sex of bovine embryos.

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      참고문헌 (Reference)

      1 "glucose-6-phosphate dehydrogenase. Theriogenology 25" 733-739,

      2 "The sexing of bovine embryos in the field." 43 : 71-90, 1995

      3 "The river buffalo (Bubalus bubalis, 2n = 50) cytogenetic map: assignment of 64 loci by fluorescence in situ hybridization and R-banding." 102 : 65-75, 2003

      4 "T. 1992. Rapid detection of chromosome aneuploidies in uncultured amniocytes by using fluorescence in situ hybridization" osatha (osatha): 55-65,

      5 "Splitting and biopsy for bovine embryo sexing under field conditions." 56 : 1383-1392, 2001

      6 "Sexing by the chromosome analysis of in vitro fertilized embryos in cattle." 20 : 179-190, 1996

      7 "Rapid sexing of preimplantation bovine embryo using consecutive and multiplex polymerase chain reaction (PCR) with biopsied single blastomere." 55 : 1843-1853, 2001

      8 "Rapid detection of male-specific DNA sequence in bovine embryos using fluorescence in situ hybridization." 51 : 390-394, 1998

      9 "R.M. 1993. Detection of aneuploid and chromosomal mosaicism in human embryos during preimplantation sex determination by fluorescent in situ hybridization" 1183-1185,

      10 "R.L. 1993. Sesitive sex determination assay applicable to bovine embryos derived from IVM and IVF. J. Reprod. Fertil. 98" 335-340,

      1 "glucose-6-phosphate dehydrogenase. Theriogenology 25" 733-739,

      2 "The sexing of bovine embryos in the field." 43 : 71-90, 1995

      3 "The river buffalo (Bubalus bubalis, 2n = 50) cytogenetic map: assignment of 64 loci by fluorescence in situ hybridization and R-banding." 102 : 65-75, 2003

      4 "T. 1992. Rapid detection of chromosome aneuploidies in uncultured amniocytes by using fluorescence in situ hybridization" osatha (osatha): 55-65,

      5 "Splitting and biopsy for bovine embryo sexing under field conditions." 56 : 1383-1392, 2001

      6 "Sexing by the chromosome analysis of in vitro fertilized embryos in cattle." 20 : 179-190, 1996

      7 "Rapid sexing of preimplantation bovine embryo using consecutive and multiplex polymerase chain reaction (PCR) with biopsied single blastomere." 55 : 1843-1853, 2001

      8 "Rapid detection of male-specific DNA sequence in bovine embryos using fluorescence in situ hybridization." 51 : 390-394, 1998

      9 "R.M. 1993. Detection of aneuploid and chromosomal mosaicism in human embryos during preimplantation sex determination by fluorescent in situ hybridization" 1183-1185,

      10 "R.L. 1993. Sesitive sex determination assay applicable to bovine embryos derived from IVM and IVF. J. Reprod. Fertil. 98" 335-340,

      11 "R.H. 1982. Survival after transfer of sexed mouse embryos exposed to H-Y antisera. Theriogenology 18" 655-662,

      12 "PCR-sexing of bovine embryos : a simplified protocol" 44 : 167-176, 1995

      13 "P.D. 1992. Mapping of repetitive bovine DNA sequences on cattle Y chromosomes. Cytogent Cell Genet. 61" 189-194,

      14 "N.L. 1993. Development of bovine embryos in vitro as affected by energy substrates. Biol. Reprod. 49" 459-462,

      15 "M.M. 1989. The use of male-specific chromosomal DNA fragments to determine the sex of bovine preimplantation embryos. Theriogenology 31" 95-104,

      16 "M. 1991. Isolation of bovine Y-derived sequence potential use in embryo sexing. Genomics 10" coti (coti): 646-653,

      17 "M. 1990. Diagnosis of the sex of bovine embryos using molecular biology. Reprod. Nutr. Dev. Suppl. 1" coti (coti): 125-132,

      18 "M. 1990. A polymorphic satellite sequence maps to the pericentric region of the bovine Y chromosome. Genomics 6" 482-490,

      19 "K.J. 1985. Production of sexed calves from frozen thawed embryos. Vet. Rec. 117" 603-608,

      20 "K.C. 1991. Micromanipulation of bovine embryos for sex determination. Theriogenology 35" 45-54,

      21 "J. 1993. A fast and efficient method for simultaneous X and Y in situ hybridization of human blastomeres. J. Assist. Reprod. Genet. 10" 82-90,

      22 "G.B. 1987. Identification of embryonic sex by detection of H-Y antigen. Theriogenology 27" 81-97,

      23 "G. 1993. Biopsy and sex determination by PCR of IVF bovine embryos. J. Reprod. Fertil. 98" 467-470,

      24 "G. 1975. Capacitation of rabbit spermatozoa in vitro. Biol. Reprod. 12" 260-274,

      25 "Frequency of sex chromosomal mosaicism in bovine embryos and its effects on sexing using a single blastomere by PCR." 11 : 87-93, 2003

      26 "E.A. 1975. Possible role of H-Y antigen in the primary determination of sex. Nature 257" 235-236,

      27 "Determining the sex of bovine embryos using polymerase chain reaction results: a six-year retrospective study." 51 : 841-854, 1999

      28 "Cryopreservation and sexing of in vivo- and in vitro-produced bovine embryos for their practical use." 50 : 29-38, 2004

      29 "Comparative analysis of Y chromosome structure in Bos taurus and B. indicus by FISH using region- specific, microdissected, and locus-specific DNA probes." 77 : 238-241, 1997

      30 "C. 1980. Transfer of cow blastocysts after sexing. Theriogenology 14" 309-318,

      31 "A. 1991. Sexing of rat embryos with antisera specific for male rats. J. Exp. Zoology 260" 99-105,

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