214 PRE-IMPLANTATION EMBRYO SEX DETERMINATION BY AMPLIFICATION OF AMELOGENIN GENE IN WATER BUFFALOES (BUBALUS BUBALIS)
J.-H. Shang A B , C.-Y. Yang A B , H.-Y. Zheng A B , J. Qin B , Y.-P. Gu A C and H.-Z. Liu A CA Key Laboratory of Buffalo Genetics, Breeding and Reproduction Technology, Ministry of Agriculture and Guangxi, Nanning, Guangxi, China;
B Buffalo Research Institute, Chinese Academy of Agricultural Sciences, Nanning, Guangxi, China;
C Faculty of Sciences, Guangdong Ocean University, Zhanjiang, Guangdong, China
Reproduction, Fertility and Development 28(2) 238-238 https://doi.org/10.1071/RDv28n2Ab214
Published: 3 December 2015
Abstract
Sex-specific sequence variability of the amelogenin (AMEL) gene has been observed in a variety of mammalian species. AMEL is a multi-copy gene expressed in abundance in the development of mammalian tooth enamel matrix. Its homologous genes are located on the X- or Y-chromosome, respectively. In our study, a pair of primers was designed to allow amplification of a single fragment of 312 bp from buffalo cows. Because the gene is slightly differently encoded on the Y-chromosome, the same primers were expected to produce 2 fragments of 312 and 249 bp from bulls. The primers were successfully applied to single, in vitro-produced Day-6 to Day-8 water buffalo blastocysts by direct lysis of the whole blastocysts, followed by PCR. The amplification on presumed female and male embryos consistently displayed the same sex-specific pattern. Of 100 blastocysts, 43 were determined to be females, 51 males, and 6 unknown (samples missed). We concluded that PCR amplification of the AMEL gene can be used for early embryo sex identification of water buffalo.
This work was supported by the National Natural Science Foundation of China under Grant No. 31160456 and the Guangxi Natural Science Foundation of China under Grants No. 2013GXNSFAA019075 and 2014GXNSFAA118116.
