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2025年8月13日 星期三
农业生物技术学报  2021, Vol. 29 Issue (3): 610-618    DOI: 10.3969/j.issn.1674-7968.2021.03.020
  研究资源与技术改进 本期目录 | 过刊浏览 | 高级检索 |
基于KASP技术的牛3种无角基因联合检测方法研究
颜泽1, 肖炜2, 张胜利1, 王雅春1, 张毅1,*
1 中国农业大学 动物科技学院,北京 100193;
2 北京市畜牧总站,北京 100107
Multiplex Detection of Three Cattle (Bos taurus) Polled Variants Based on KASP Assay
YAN Ze1, XIAO Wei2, ZHANG Sheng-Li1, WANG Ya-Chun1, ZHANG Yi1,*
1 College of Animal Science and Technology, China Agricultural University, Beijing 100193, China;
2 Beijing General Station of Animal Husbandry, Beijing 100107, China
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摘要 在自然界,角是野生动物抵御天敌、争夺配偶的工具,但在规模化养殖中,天生无角是有利性状,可减少管理成本和保护动物福利。牛(Bos taurus)无角性状呈常染色体显性遗传,具有多等位基因特点,目前已知有3类不同来源的无角等位基因(奶牛: Friesian; 肉牛、乳肉兼用牛: Celtic; 蒙古牛: Mongolian)。为了开发准确高效的牛无角基因分子检测方法,研究基于整合竞争性等位基因特异性PCR (kompetitive allele-specific PCR, KASP)和微流控芯片技术的检测平台,根据3种不同类型的结构变异设计特异性引物,建立了针对3种牛无角基因的快速诊断方法。通过对采自国内外不同牛品种的样本检测,并与PCR扩增测序和凝胶电泳等传统检测方法的结果相对比,证明新方法可以对3种无角基因准确分型,适于规模化检测应用。此外,本研究首次在我国三河牛品种中鉴定出Mongolian无角等位基因,同时在Friesian无角等位基因序列中新发现一个2 bp缺失,可作为无角基因特异标记。研究结果为开展无角牛的分子选育提供了技术手段。
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颜泽
肖炜
张胜利
王雅春
张毅
关键词 竞争性等位基因特异性PCR (KASP)无角基因三河牛分子检测微流控芯片    
Abstract:In wild animals, horns are used to fight for survival and mates. However, in the intensive farming system, polledness is a favorable trait because helps avoid the cost and protect animal welfare. Polledness is an autosomal dominant trait with multiple variants/alleles. Three types of polled variants have been reported on chromosome 1 of taurine cattle (Bos taurus) - the Friesian variant that occurred in dairy cattle breeds, the Celtic variant that was found in beef and dual-purposed breeds, and the Mongolian variant that was identified in Mongolian cattle. In this study, based on a genotyping platform that combines KASP (kompetitive allele-specific PCR) assay and microfluidic chip, a molecular diagnostic method was developed to detect the three polled variants simultaneously. The new KASP assay was further validated by using the conventional molecular experiments including PCR amplification, gel electrophoresis and Sanger sequencing, demonstrating that the new method was accurate and reliable and could be applied in large-scale testing. Interestingly, the Mongolian polled variant was detected in a bull of Sanhe cattle, an indigenous Chinese cattle breed. In addition, a previously unpublished deletion of 2 bp was identified in the Friesian variant that could be used as a marker associated with polledness. These results provide a technical basis for molecular breeding for polled cattle.
Key wordsKompetitive allele-specific PCR (KASP)    Polled gene    Sanhe cattle    Molecular assay    Microfluidic chip
收稿日期: 2020-08-04      出版日期: 2021-03-01
ZTFLH:  S823  
基金资助:现代农业(奶牛)产业技术体系建设专项资金(CARS-36); 北京市奶牛产业创新团队项目(BAIC06)
通讯作者: *yizhang@cau.edu.cn   
引用本文:   
颜泽, 肖炜, 张胜利, 王雅春, 张毅. 基于KASP技术的牛3种无角基因联合检测方法研究[J]. 农业生物技术学报, 2021, 29(3): 610-618.
YAN Ze, XIAO Wei, ZHANG Sheng-Li, WANG Ya-Chun, ZHANG Yi. Multiplex Detection of Three Cattle (Bos taurus) Polled Variants Based on KASP Assay. 农业生物技术学报, 2021, 29(3): 610-618.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2021.03.020     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2021/V29/I3/610
 
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