采用四引物ARMS-PCR方法检测奶牛乳腺组织DGAT1基因单核苷酸多态性

发布时间：2018-02-14 16:34

本文关键词： 荷斯坦奶牛 DGAT基因 ARMS-PCR 基因分型　出处：《中国畜牧兽医》2017年10期 　论文类型：期刊论文

【摘要】：试验旨在快速有效地检测泌乳奶牛的二酰甘油转酰基酶1(diacylgycerol acyltransferase 1,DGAT1)乳脂量性状的优势等位基因K232A单核苷酸多态性,确定DGAT1基因型进而确定泌乳性状,为中国荷斯坦奶牛分子标记辅助选择提供技术支持。选取6头泌乳初期(3头为高乳产量牛,3头为低乳产量牛)中国北方荷斯坦奶牛为研究对象,提取乳腺组织基因组DNA,分别设计1对外引物和1对内引物,建立一种四引物ARMS-PCR体系快速检测奶牛乳腺组织DGAT1基因单核苷酸多态性。结果发现,外引物扩增片段长度为512 bp,为PCR反应的阳性对照,基因型为232K扩增片段长度为369 bp,基因型为232A扩增片段长度为181 bp。PCR结果显示,6头牛的乳腺组织样本均由外部引物扩增出长度为512 bp的片段,泌乳期高乳产量奶牛和泌乳期低乳产量奶牛乳腺组织的特异性扩增片段长度均为181 bp。表明本研究选取的6头奶牛样本DGAT1基因K232A多态性均为232A型。提示该PCR鉴定方法能够快速有效地鉴定奶牛DGAT1基因型,可用于中国荷斯坦奶牛分子标记辅助选择。
[Abstract]:The aim of the experiment was to detect the dominant allele K232A single nucleotide polymorphism of diacylglycerol transacylgycerol acyltransferase 1 DGAT1 in lactating cows quickly and effectively, and to determine the DGAT1 genotype and then the lactation traits. To provide technical support for molecular marker-assisted selection of Chinese Holstein cows, 6 Holstein cows with high milk yield and 3 low milk yield cows were selected as the research objects. The genomic DNA of mammary gland tissue was extracted, 1 external primer and 1 pair of internal primer were designed, and a four primer ARMS-PCR system was established for rapid detection of single nucleotide polymorphism of DGAT1 gene in dairy cow mammary tissue. The amplified fragment length of outer primer was 512bp, which was the positive control of PCR reaction. The length of the amplified fragment was 369bp for genotype 232K and 181 bp.PCR for genotype 232A. The results showed that 512bp fragment was amplified from breast tissue samples of 6 cattle by external primer. The specific amplified fragment length of breast tissue of high lactating dairy cows and low lactating dairy cows was 181 BP. The results showed that the K232A polymorphism of DGAT1 gene was 232A type in 6 dairy cattle samples selected in this study. It is suggested that this PCR identification formula is suitable for this purpose. The method can be used to identify the DGAT1 genotypes of dairy cows quickly and effectively. It can be used for molecular marker-assisted selection in Chinese Holstein cows.
【作者单位】：东北农业大学生命科学学院;
【基金】：国家自然科学基金面上项目(31671285) 黑龙江省博士后科研启动金(LBH-Q16027)
【分类号】：S823

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