奶牛乳脂性状候选基因EEF1D突变位点功能分析

doi:10.3969/j.issn.1004-1524.2020.07.03

浙江农业学报 ›› 2020, Vol. 32 ›› Issue (7): 1155-1159.DOI: 10.3969/j.issn.1004-1524.2020.07.03

奶牛乳脂性状候选基因EEF1D突变位点功能分析

蒋秋斐^1,2, 蔡正云¹, 黄增文¹, 冯小芳¹, 张娟¹, 顾亚玲^1,*

1.宁夏大学农学院,宁夏银川 750021;
2.宁夏回族自治区畜牧工作站,宁夏银川 750004

收稿日期:2019-07-29 出版日期:2020-07-25 发布日期:2020-07-28
通讯作者: *顾亚玲,E-mail:guyaling@sina.com
作者简介:蒋秋斐(1987—),女,宁夏银川人,博士研究生,畜牧师,研究方向为动物遗传育种与繁殖。E-mail:jiangqiufei@163.com
基金资助:
宁夏大学科学研究基金(ZR1725); 宁夏回族自治区农业育种专项(2019NYYZ05)

Functional analysis of EEF1D mutation site in dairy cow milk fat traits candidate gene

JIANG Qiufei^1,2, CAI Zhengyun¹, HUANG Zengwen¹, FENG Xiaofang¹, ZHANG Juan¹, GU Yaling^1,*

1. School of Agriculture, Ningxia University, Yinchuan 750021, China;
2. Animal Husbandry Station of Ningxia, Yinchuan 750004, China

Received:2019-07-29 Online:2020-07-25 Published:2020-07-28

摘要/Abstract

摘要： 提高乳脂含量、提升奶质已成为当前奶牛业研究的重点方向之一。前期经GWAS方法研究发现,EEF1D基因5'非翻译区(5'-UTR)的G/A突变与乳脂率极显著相关,为了进一步对候选基因EEF1D的突变位点进行功能分析,首先通过人工合成获得了EEF1D基因突变型与野生型片段,并在细胞水平上对其开展了功能验证实验。研究发现,EEF1D基因5'-UTR的G/A突变引起Sp3转录起始位点后移33 bp,并且突变型的活性大约是野生型的3倍。研究结果表明,该突变可改变转录因子的转录起始位点,从而使EEF1D的表达上调。

关键词: 乳脂含量, EEF1D基因, 5'非翻译区(5'-UTR), Sp3, 突变型

Abstract: Increasing milk fat content and improving milk quality have become one of the key research directions in the dairy industry. In the previous study, the G/A mutation in the 5'-untranslated region (5'-UTR) of EEF1D gene was significantly correlated with the milk fat percentage. In order to further analyze the functional site of the candidate gene EEF1D, the EEF1D gene mutant and wild-type fragments were obtained by artificial synthesis, and functional verification experiments were carried out at the cellular level. This study found that the G/A mutation of the 5'-UTR of the EEF1D gene caused a 33 bp shift in the Sp3 transcription initiation site, and the activity of the mutant was about three times that of the wild type. These findings indicated that this mutation can alter the transcription initiation site of the transcription factor, thereby up-regulating the expression of EEF1D.

Key words: milk fat content, EEF1D gene, 5' untranslated region (5'-UTR), Sp3, mutant

中图分类号:

S823.9⁺1

蒋秋斐, 蔡正云, 黄增文, 冯小芳, 张娟, 顾亚玲. 奶牛乳脂性状候选基因EEF1D突变位点功能分析[J]. 浙江农业学报, 2020, 32(7): 1155-1159.

JIANG Qiufei, CAI Zhengyun, HUANG Zengwen, FENG Xiaofang, ZHANG Juan, GU Yaling. Functional analysis of EEF1D mutation site in dairy cow milk fat traits candidate gene[J]. , 2020, 32(7): 1155-1159.

参考文献

[1] MULNER-LORILLON O, MINELLA O, CORMIER P, et al.Elongation factor EF1d, a new target for maturationpromoting factor in Xenopus oocytes[J]. Journal of Biological Chemistry, 1994, 269(31):20201-20207.
[2] GYENIS L, DUNCAN J S, TUROWEC J P, et al.Unbiased functional proteomics strategy for protein kinase inhibitor validation and identification of bona fide protein kinase substrates: application to identification of EEF1D as a substrate for CK2[J]. Journal of Proteome Research, 2011, 10(11): 4887-4901.
[3] DELALANDE C, MONNIER A, MINELLA O, et al.Developmental regulation of elongation factor-1 δ in Sea urchin suggests appearance of a mechanism for alternative poly(A) site selection in gastrulae[J]. Experimental Cell Research, 1998, 242(1): 228-234.
[4] 谢岩. 奶牛产奶性状候选基因EEF1D和PDE9A功能验证[D]. 北京: 中国农业大学, 2013.
XIE Y.Identification and functional verification of EEF1D and PDE9A associated with milk production traits in dairy cattle[D]. Beijing: China Agricultural University, 2013.(in Chinese with English abstract)
[5] CHEN F C, PAN C L, LIN H Y. Functional implications of RNA splicing for human long intergenic noncoding RNAs[J]. Evolutionary Bioinformatics, 2014, 10: EBO.S20772.
[6] CHEN L, BUSH S J, TOVAR-CORONA J M, et al. Correcting for differential transcript coverage reveals a strong relationship between alternative splicing and organism complexity[J]. Molecular Biology and Evolution, 2014, 31(6): 1402-1413.
[7] WANG Q L, ZHOU T S.Alternative-splicing-mediated gene expression[J]. Physical Review E, 2014, 89: 012713.
[8] RAVI S, SCHILDER R J, KIMBALL S R.Role of precursor mRNA splicing in nutrient-induced alterations in gene expression and metabolism[J]. The Journal of Nutrition, 2015, 145(5): 841-846.
[9] ZHANG S H, WU X F, PAN C Y, et al.Identification of novel isoforms of dairy goat EEF1D and their mRNA expression characterization[J]. Gene, 2016, 581(1): 14-20.
[10] 刘轩. 基于目标区域捕获测序策略进行奶牛产奶性状功能基因挖掘[D]. 北京: 中国农业大学, 2014.
LIU X.Targeted resequencing of GWAS loci reveals novel functional genes for milk production traits[D]. Beijing: China Agricultural University, 2014.(in Chinese with English abstract)
[11] 龙小娟. 牛AGPAT6基因遗传多态性研究及与脂代谢相关功能的验证[D]. 长春: 吉林大学, 2016.
LONG X J.Polymorphism detection and function verifiction with lipid metabolism of AGPAT6 gene in cattle[D]. Changchun: Jilin University, 2016.(in Chinese with English abstract)
[12] TENAILLON M I, SAWKINS M C, LONG A D, et al.Patterns of DNA sequence polymorphism along chromosome 1 of maize (Zea mays ssp. mays L.)[J]. Proceedings of the National Academy of Sciences of the United States of America, 2001, 98(16): 9161-9166.
[13] ASLAM M L, BASTIAANSEN J W, CROOIJMANS R P, et al.A SNP based linkage map of the turkey genome reveals multiple intrachromosomal rearrangements between the turkey and chicken genomes[J]. BMC Genomics, 2010, 11(1): 647.
[14] BARENDSE W, HARRISON B E, BUNCH R J, et al.Genome wide signatures of positive selection: the comparison of independent samples and the identification of regions associated to traits[J]. BMC Genomics, 2009, 10(1): 178.
[15] OKAMOTO A, TAINAKA K, OCHI Y, et al.Simple SNP typing assay using a base-discriminating fluorescent probe[J]. Molecular BioSystems, 2006, 2(2): 122.
[16] ELENITOBA-JOHNSON K S J, BOHLING S D. Solution-based scanning for single-base alterations using a double-stranded DNA binding dye and fluorescence-melting profiles[J]. The American Journal of Pathology, 2001, 159(3): 845-853.
[17] RAFALSKI A.Applications of single nucleotide polymorphisms in crop genetics[J]. Current Opinion in Plant Biology, 2002, 5(2): 94-100.
[18] SUSKE G, BRUFORD E, PHILIPSEN S.Mammalian SP/KLF transcription factors: bring in the family[J]. Genomics, 2005, 85(5): 551-556.

奶牛乳脂性状候选基因EEF1D突变位点功能分析

Functional analysis of EEF1D mutation site in dairy cow milk fat traits candidate gene

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