Sex determination of porcine embryos using a new developed duplex polymerase chain reaction procedure based on the amplification of repetitive sequences
作者： Eva Torner , Eva Bussalleu , M. Dolors Briz , Alfonso Gutiérrez-Adán , Sergi Bonet
DOI: 10.1071/RD12033
关键词:
摘要: Polymerase chain reaction (PCR)-based assays have become increasingly prevalent for sexing embryos. The aim of the present study was to develop a suitable duplex PCR procedure based on amplification porcine repetitive sequences tissues, embryos and single cells. Primers were designed targeting X12696 Y chromosome-specific repeat sequence (SUSYa SUSYb; sex-related primer sets), multicopy porcine-specific mitochondrial 12S rRNA gene (SUS12S; control set) X51555 1 chromosome (SUS1; set). specificity sets established technique optimised by testing combinations two specific (SUSYa/SUS12S; SUSYb/SUS12S), different concentrations, sources DNA polymerase, melting temperatures numbers cycles using genomic from ovarian testicular tissue. SUSYa/SUS12S- SUSYb/SUS12S-based procedures applied in vitro-produced (IVP) blastocysts, cell-stage oocytes. SUSYb/SUS12S primer-based successfully sexed cells IVP (100% efficiency), as well blastocysts (96.6% accuracy; 96.7% efficiency). This is first report demonstrate applicability these this purpose. In conclusion, highly reliable sensitive
sci-hub.se 参考文章(51)
Johnson La, Advances in gender preselection in swine. Journal of reproduction and fertility. ,vol. 52, pp. 255- 266 ,(1997)
D Rath, C R Long, J R Dobrinsky, G R Welch, L L Schreier, L A Johnson, In vitro production of sexed embryos for gender preselection: high-speed sorting of X-chromosome-bearing sperm to produce pigs after embryo transfer. Journal of Animal Science. ,vol. 77, pp. 3346- 3352 ,(1999) , 10.2527/1999.77123346X
Steve Rozen, Helen Skaletsky, Primer3 on the WWW for general users and for biologist programmers. Methods of Molecular Biology. ,vol. 132, pp. 365- 386 ,(2000) , 10.1385/1-59259-192-2:365
D. Pomp, B. A. Good, R. D. Geisert, C. J. Corbin, A. J. Conley, Sex identification in mammals with polymerase chain reaction and its use to examine sex effects on diameter of day-10 or -11 pig embryos. Journal of Animal Science. ,vol. 73, pp. 1408- 1415 ,(1995) , 10.2527/1995.7351408X
B.P. Chowdhary, T. Raudsepp, The horse genome. Genome dynamics. ,vol. 2, pp. 97- 110 ,(2006) , 10.1159/000095098
J.H. Park, J.H. Lee, K.M. Choi, S.Y. Joung, J.Y. Kim, G.M. Chung, D.I. Jin, K.S. Im, Rapid sexing of preimplantation bovine embryo using consecutive and multiplex polymerase chain reaction (PCR) with biopsied single blastomere. Theriogenology. ,vol. 55, pp. 1843- 1853 ,(2001) , 10.1016/S0093-691X(01)00526-X
D.L. Garner, G.E. Seidel, History of commercializing sexed semen for cattle Theriogenology. ,vol. 69, pp. 886- 895 ,(2008) , 10.1016/J.THERIOGENOLOGY.2008.01.006
Anne R. Greenlee, Rebecca L. Krisher, Edward D. Plotka, Rapid sexing of murine preimplantation embryos using a nested, multiplex polymerase chain reaction (PCR). Molecular Reproduction and Development. ,vol. 49, pp. 261- 267 ,(1998) , 10.1002/(SICI)1098-2795(199803)49:3<261::AID-MRD6>3.0.CO;2-M
P. Bredbacka, Progress on methods of gene detection in preimplantation embryos. Theriogenology. ,vol. 55, pp. 23- 34 ,(2001) , 10.1016/S0093-691X(00)00443-X
Atsushi Akane, Satoko Seki, Hiroshi Shiono, Hiroaki Nakamura, Masanori Hasegawa, Masato Kagawa, Kazuo Matsubara, Yutaka Nakahori, Shigeo Nagafuchi, Yasuo Nakagome, Sex determination of forensic samples by dual PCR amplification of an X-Y homologous gene Forensic Science International. ,vol. 52, pp. 143- 148 ,(1992) , 10.1016/0379-0738(92)90102-3