Abstract
Insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2, also called IMP2) plays an essential role in the development and maturation of germ cells and embryos and is a candidate gene for goat litter size, based on a previous genome-wide selective sweep analysis. In this study, the mRNA expression level of IGF2BP2 was found to be significantly higher in a single-lamb group than in a multi-lamb group. Insertions/deletions (indels) within the goat IGF2BP2 gene, including P4-Ins-13bp and P5-Del-12bp, were verified in 918 Shaanbei White Cashmere (SBWC) female goats. The minor allelic frequencies (MAFs) of P4-Ins-13bp and P5-Del-12bp loci were 0.349 and 0.295, respectively. Analysis using the Chi-square (χ2) test showed that the genotype (χ2=14.479, P=0.006) distribution of P4-Ins-13bp was significantly different between the single-lamb and multi-lamb groups. Correlation analysis demonstrated that P4-Ins-13bp was significantly associated with goat litter size (P=0.022), and individual goats with the homozygous deletion/deletion (DD) genotype produced more litters than other goats. Therefore, considered as a potential molecular marker significantly related to lambing traits, the P4-Ins-13bp mutation of the goat IGF2BP2 gene can be used in goat breeding with practical molecular marker-assisted selection (MAS) to optimize female reproduction and improve economic efficiency in the goat industry.
概要
目的
验证IGF2BP2插入/缺失与陕北白绒山羊产羔数之间的关系, 并选择有利的等位基因.
创新点
首次筛选、 验证山羊IGF2BP2基因的InDel位点, 确定其与山羊产羔性状的相关性及作用, 为山羊经济性状改良及新品种培育提供理论依据.
方法
采用实时荧光定量聚合酶链式反应(qRT-PCR)方法分析IGF2BP2在单羔组和多羔组中的表达水平; 通过PCR扩增-琼脂糖凝胶电泳在918只陕北白绒母山羊中鉴定IGF2BP2基因P4-Ins-13bp和P5-Del-12bp缺失/插入位点基因型, 并进行测序以验证突变; 利用SHEsis平台分析两个位点的连锁不平衡; 采用方差分析(ANOVA)和χ2检验分析山羊IGF2BP2基因变异与产羔数的关系; 并利用在线数据库预测转录因子(TF)和突变区域序列的结合.
结论
本研究发现, IGF2BP2的mRNA表达水平在单羔组明显高于多羔组. 在陕北白绒母山羊中验证了IGF2BP2基因内的插入/缺失(indels), 包括P4-Ins-13bp和P5-Del-12bp. χ2检验表明, P4-Ins-13bp基因型(χ2=14.479, P=0.006)在单羔和多羔组间分布差异显著; 相关分析表明, P4-Ins-13bp与山羊产仔数显着相关(P=0.022), 具有缺失/缺失(DD)基因型的山羊个体产仔数高于其他山羊. 因此, IGF2BP2基因的P4-Ins-13bp突变可作为一种潜在的与繁殖性状显著相关的分子标记, 以优化母羊繁殖力, 提高山羊产业的经济效益.
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Acknowledgments
This study was supported by the National Natural Science Foundation of China (Nos. 32060734 and 31760650).
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Contributions
Xianyong LAN and Dongyun XIN designed the experiment; Haijing ZHU, Lei QU, and Libang HE provided goat samples and data records; Dongyun XIN and Yangyang BAI carried out all the experiments; Dongyun XIN prepared all figures and tables and drafted the manuscript; Yi BI, Yuxin KANG, Hong CHEN, Xianyong LAN, and Chuanying PAN checked and revised the manuscript. All authors have read and approved the final manuscript and, therefore, have full access to all the data in the study and take responsibility for the integrity and security of the data.
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Dongyun XIN, Yangyang BAI, Yi BI, Libang HE, Yuxin KANG, Chuanying PAN, Haijing ZHU, Hong CHEN, Lei QU, and Xianyong LAN declare that they have no conflict of interest.
The animal participants involved in this experiment were maintained in strict accordance with the “Regulations on the Management of Experimental Animal Affairs” (Ministry of Science and Technology of China, 2004) and this study was approved by the International Animal Care and Use Committee (IACUC) of Northwest A&F University, Yangling, China (Protocol No. NWAFAC1008).
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Xin, D., Bai, Y., Bi, Y. et al. Insertion/deletion variants within the IGF2BP2 gene identified in reported genome-wide selective sweep analysis reveal a correlation with goat litter size. J. Zhejiang Univ. Sci. B 22, 757–766 (2021). https://doi.org/10.1631/jzus.B2100079
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DOI: https://doi.org/10.1631/jzus.B2100079
Key words
- Goat
- Insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2)
- Litter size
- Indel
- Marker-assisted selection (MAS)