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首页> 外文期刊>Molecular reproduction and development >Sex determination in cattle based on simultaneous amplification of a new male-specific DNA sequence and an autosomal locus using the same primers.
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Sex determination in cattle based on simultaneous amplification of a new male-specific DNA sequence and an autosomal locus using the same primers.

机译：基于使用相同引物同时扩增新的男性特异性DNA序列和常染色体基因座，确定牛的性别。

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摘要

A PCR-based method for sex determination of bovine DNA samples and embryo biopsies is presented. Using only one primer pair both the male-specific sequence FBNY (127 bp) and a sex-independent control PCR-fragment, the microsatellite marker FBN17 (136-140 bp) are generated in the same PCR reaction. Synteny mapping assigned the male-specific sequence to bovine chromosome Y (BTA Y), whereas FBN17 was mapped to bovine chromosome 2. Localisation of FBNY on BTA Y was confirmed by fluorescence in hybridisation of two BAC clones containing the male-specific sequence. There was no amplification of the male-specific target sequence FBNY in sheep, pig, goat, mice, man, and several wild species of the tribe Bovini. The bovine male-specific fragment was detected in dilutions containing as little as 10 pg genomic DNA and in blastomeres from embryo biopsies. The PCR assay presented here does require neither restriction endonuclease digestion of the PCR product nor additional nested PCR steps. Owing to the advantage of parallel amplification of the autosomal locus FBN17 no additional control fragment is necessary to detect PCR failure. The results of sex determination in embryo biopsies using FBNY were in agreement with the outcome from a reference assay used in commercial breeding programs. Mol. Reprod. Dev. 60: 13-19, 2001. Copyright 2001 Wiley-Liss, Inc.
机译：提出了一种基于PCR的牛DNA样品和胚胎活检性别确定方法。仅使用一对引物，雄性特异性序列FBNY（127 bp）和性别无关的对照PCR片段均在同一PCR反应中生成了微卫星标记FBN17（136-140 bp）。通过序列映射将雄性特异性序列分配给牛Y染色体（BTA Y），而将FBN17映射到牛染色体2。通过在两个包含雄性特异性序列的BAC克隆杂交中的荧光证实了FBNY在BTA Y上的定位。在绵羊，猪，山羊，小鼠，人和博维尼部落的几个野生物种中，男性特异性靶序列FBNY没有扩增。在含有低至10 pg基因组DNA的稀释液和胚胎活检的卵裂球中检测到了牛雄性特异性片段。此处介绍的PCR测定法确实不需要PCR产物的限制性核酸内切酶消化，也不需要额外的嵌套PCR步骤。由于平行扩增常染色体基因座FBN17的优势，无需其他对照片段即可检测PCR失败。使用FBNY在胚胎活检中确定性别的结果与商业育种计划中使用的参考测定的结果一致。大声笑责备。开发人员60：13-19，2001。版权所有2001 Wiley-Liss，Inc.。

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