Hostname: page-component-586b7cd67f-dlnhk Total loading time: 0 Render date: 2024-11-23T05:36:51.140Z Has data issue: false hasContentIssue false

Establishing and optimizing the system for sex determination of bovine preimplantation embryos

Published online by Cambridge University Press:  12 February 2007

Chen Cong-Ying
Affiliation:
Key Laboratory for Animal Biotechnology of Jiangxi Province and the Ministry of Agriculture, Jiangxi Agricultural University, Nanchang, 330045, China
Huang Lu-Sheng*
Affiliation:
Key Laboratory for Animal Biotechnology of Jiangxi Province and the Ministry of Agriculture, Jiangxi Agricultural University, Nanchang, 330045, China
Chen Jing-Bo
Affiliation:
Institute of Animal Science, Xinjiang Academy of Animal Science, Urumqi, 830000, China
Ding Neng-Shui
Affiliation:
Key Laboratory for Animal Biotechnology of Jiangxi Province and the Ministry of Agriculture, Jiangxi Agricultural University, Nanchang, 330045, China
Zhou Li-Lua
Affiliation:
Key Laboratory for Animal Biotechnology of Jiangxi Province and the Ministry of Agriculture, Jiangxi Agricultural University, Nanchang, 330045, China
Xiao Hai-Xia
Affiliation:
Institute of Animal Science, Xinjiang Academy of Animal Science, Urumqi, 830000, China
*
*Corresponding author. E-mail: [email protected]

Abstract

Two pairs of nested primers were derived from the 5′ flanking sequence of the bovine SRY gene to serve as male-specific sexing primers, which were used for PCR-based sex determination of bovine embryos in combination with casein protein gene primers as internal control. At the same time, methods of DNA extraction from bovine embryo cells were established and the sensitivity of nested PCR and normal PCR was studied for developing a technique of sex determination of bovine embryos adapted for practical use. Results showed that the primers used in the study were male-specific and all primers were bovine-specific. Boiling and freeze–thawing were used for DNA extraction from embryo cells. Amplification products were obtained with only 10 cells by nested PCR, so this system was used for the sex determination of bovine embryos. The sex of 10 Holstein cow embryos was identified and the results were confirmed upon birth of the calves.

Type
Research Article
Copyright
Copyright © China Agricultural University and Cambridge University Press 2004

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Aasen, E and Medrano, JF (1990) Amplification of the ZFY and ZFX genes for sex identification in humans, cattle, sheep and goats. Biotechnology 8: 12791281.Google ScholarPubMed
Agrawala, PL, Wagner, VA and Geldermann, H (1992) Sex determination and milk protein genotyping of preimplantation stage bovine embryos using multiplex PCR. Theriogenology 38: 969978.CrossRefGoogle ScholarPubMed
Appa Rao, KB and Totey, SM (1999) Cloning and sequencing of buffalo male-specific repetitive DNA: sexing of in-vitro developed buffalo embryos using multiplex and nested polymerase chain reaction. Theriogenology 51: 785797.CrossRefGoogle ScholarPubMed
Huang, S-Z, Chen, M-J, Huang, Y et al. ,(2000) Sexing the IVF bovine and goat embryos. Hereditas 22: 6568. in Chinese with English abstractGoogle Scholar
Park, JH, Lee, JH, Choi, KM et al. ,(2001) Rapid sexing of preimplantation bovine embryo using consecutive and multiplex polymerase chain reaction (PCR) with biopsied single blastomere. Theriogenology 55: 18431853.CrossRefGoogle ScholarPubMed
Sambrook, J, Fritch, EF, Maniatis, T et al. ,(1989) Molecular Cloning: A Laboratory Manual, 2nd ed. New York: Cold Spring Harbor Laboratory Press.Google Scholar
Schroder, A, Miller, J, Thomsen, P et al. ,(1991) Sex determination of bovine embryos using the polymerase chain reaction. Animal Biotechnology 1: 121133.CrossRefGoogle Scholar
Yuko, K, Seijisato Xiao, C et al. ,(1995) Cloning and characterization of bovine SRY gene. Animal Science and Technology 66: 9941001.Google Scholar
Zeng, Y-T, Ren, Z-R, Hu, M-X et al. , (1993) Sexing cow embryos by PCR amplification of SRY sequence. Science in China. Series B, Chemistry, Life Sciences & Earth Sciences 23: 371375 (in Chinese).Google Scholar