摘要
大量研究发现赛鸽中LDHA、F-KER、DRD4、MSTN和CRY1这5个基因存在单核苷酸多态性,并与归巢能力相关,成为分子辅助育种的潜在SNP标记。当前的检测方法仍以传统的RFLP为主,尚未报道Sanger测序的应用。通过建立一种基于Sanger测序技术的检测方法,以识别这5个基因中的多态性位点。结果表明:针对赛鸽的LDHA、F-KER、DRD4、MSTN和CRY1基因设计了特异性引物,用于扩增单核苷酸多态性位点,所得PCR产物条带单一,显示具有良好的特异性。进一步,利用这些引物对60羽赛鸽的羽毛DNA样品进行扩增和Sanger测序,能成功获得各个基因位点的多态性分型结果。最后,统计了各种基因分型在这60羽赛鸽中的分布频率,发现相关的罕见基因型频率均低于5%,与国外赛鸽群体的频率相一致。研究显示所建立的赛鸽竞翔相关基因SNP分型Sanger测序分析方法,对于选育优良基因型的赛鸽具有重要的应用价值。
A large number of studies have found that the five genes of LDHA,F-KER,DRD4,MSTN and CRY1 in the racing pigeons had single nucleotide polymorphisms,which were associated with the homing ability,making them become the potential SNP markers for molecular assisted breeding.The current detection methods were still based on the traditional RFLP,and the application of Sanger sequencing has not been reported.A detection method based on Sanger sequencing technology was established to identify the polymorphic sites in these five genes.The results showed that:the specific primers were designed for LDHA,F-KER,DRD4,MSTN and CRY1 genes in the racing pigeons to amplify the single nucleotide polymorphism sites.The obtained PCR products had single bands,showing good specificity.Further,the amplification and Sanger sequencing were performed on the feather DNA samples from 60 racing pigeons by using these primers.And the polymorphic genotyping results for each gene locus were successfully obtained.Finally,the distribution frequency of various genotypes among these 60 racing pigeons was statistically analyzed,and it was found that the frequencies of the relevant rare genotypes were all less than 5%,which were consistent with the frequencies reported in the foreign racing pigeon populations.The study showed that the established Sanger sequencing analysis method for SNP genotyping of racing pigeon performance-related genes had important application value for the breeding of racing pigeons with excellent genotypes.
作者
杨理程
吴锐琼
李培健
王冰梅
黄镇
YANG Li-cheng;WU Rui-qiong;LI Pei-jian;WANG Bing-mei;HUANG Zhen(College of Life Sciences,Fujian Normal University,Fuzhou,Fujian 350117,China;Fujian Strait Pigeon Culture Media Co.,Ltd.,Fuzhou,Fujian 350000,China)
出处
《福建农业科技》
CAS
2024年第7期1-9,共9页
Fujian Agricultural Science and Technology
基金
福建省自然科学基金项目(2022J01637)。
关键词
赛鸽
Sanger测序法
基因分型
Racing pigeons
Sanger sequencing method
Genetic typing
