牦牛胚胎性别鉴定PCR反应体系的建立

畜牧兽医学报 ›› 2011, Vol. 42 ›› Issue (1): 141-144.doi:

牦牛胚胎性别鉴定PCR反应体系的建立

张大伟，字向东*，黄磊，马力，陈达文，徐华伟,梁冠男

西南民族大学高原动物遗传育种与繁殖教育部重点实验室，成都 610041

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-01-25 发布日期:2011-01-25
通讯作者: 字向东

Sexing of in vitro-produced Yak Embryos by Nested-PCR

ZHANG Da-wei, ZI Xiang-dong*, HUANG Lei, MA Li, CHEN Da-wen, XU Hua-wei, LIANG Guan-nan

The Key Laboratory of Plateau Animal Genetics, Breeding and Reproduction, Ministry of Education, Southwest University for Nationalities, Chengdu 610041, China

Received:1900-01-01 Revised:1900-01-01 Online:2011-01-25 Published:2011-01-25

摘要/Abstract

摘要： 研究旨在建立牦牛胚胎性别鉴定准确、可靠的PCR反应体系及研究切割取样对胚胎发育能力的影响。根据牦牛SRY和HSL基因序列分别设计合成2对巢式PCR引物作为性别鉴定引物和2对引物作为内标引物，通过优化PCR反应条件，对已知性别牦牛血液基因组DNA和牦牛杂种胚胎DNA采用巢式PCR进行性别鉴定。结果表明，血液样本性别鉴定与实际性别完全相符，准确率100%。从早期胚胎取8个细胞进行性别鉴定，结果雄性为45.8%（11/24），雌性为54.2%(13/24)，取样后胚胎发育率为56.7%。结果说明已成功建立牦牛胚胎性别鉴定的PCR体系。

Abstract: The objective of this study was to obtain an accurate and reliable method for determining the sex of yak embryos through amplification of SRY gene of the yaks, and evaluate embryonic development after biopsies. Two pairs of SRY gene nested primers for sex determination and two pairs of HSL gene primers as internal standard and optimized PCR reaction conditions were designed. The accuracy of PCR amplification for sex determination was verified by 24 blood samples. The result showed that in every case, the results were as expected for both female and male, i.e., accuracy was 100%. Using this optimized procedure, clear signals following PCR amplification were obtained from all samples of IVFderived yak embryos, eight cells were sampled from each embryo, 45.8% (11/24) embryos for males and 54.2% (13/24) for females. 56.7% (17/30) of biopsied embryos could further develop. It was concluded that this PCR system was highly reliable for sex determination of yak embryos.

张大伟;字向东;黄磊;马力;陈达文;徐华伟;梁冠男 . 牦牛胚胎性别鉴定PCR反应体系的建立[J]. 畜牧兽医学报, 2011, 42(1): 141-144.

ZHANG Da-wei;ZI Xiang-dong;HUANG Lei;MA Li;CHEN Da-wen;XU Hua-wei;LIANG Guan-nan . Sexing of in vitro-produced Yak Embryos by Nested-PCR[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2011, 42(1): 141-144.

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