波尔山羊PLAG1基因克隆、多态性鉴定及其与出生体重、体尺性状的关联分析

doi:10.16431/j.cnki.1671-7236.2022.09.020

摘要/Abstract

摘要： 【目的】试验旨在研究多形性腺瘤基因1（plemorphic adenoma gene 1，PLAG1）基因序列特征、多态性以及对山羊出生体重、体尺的影响。【方法】以波尔山羊为研究对象，克隆PLAG1基因序列并测序，采用实时荧光定量PCR鉴定其在不同年龄和体重波尔山羊组织中的表达模式，采用Western blotting方法检测PLAG1在不同体重羔羊腿肌中的表达水平，进一步筛选PLAG1基因CDS和3'-UTR区SNP，并分析各位点不同基因型与波尔山羊出生体重、体尺的相关性。【结果】波尔山羊PLAG1基因CDS全长1 500 bp，编码499个氨基酸，PLAG1蛋白相对保守。组织表达谱显示，PLAG1基因在出生体重较大的羔羊心脏、小肠和腿肌中表达水平显著或极显著高于出生体重较小的羔羊，在胎羊各组织中mRNA表达水平均显著或极显著高于成年母羊（P<0.05；P<0.01）；PLAG1蛋白在出生体重较大的羔羊腿肌中表达量极显著高于出生体重较小的羔羊（P<0.01）。在波尔山羊PLAG1基因3'-UTR区共筛选到6个SNPs位点，分别为：2664 A>G、2712 A>G、2721 T>C、2879 T>A、2990 A>T和3270 A>G。关联分析发现，2664 A>G位点AG基因型个体出生管围显著大于GG基因型（P<0.05）；2721 T>C位点TT基因型个体出生体长显著大于TC基因型（P<0.05）；2879 T>A位点TA基因型个体出生管围显著大于AA基因型（P<0.05）；2990 A>T位点AA基因型个体出生体重显著高于AT基因型（P<0.05）；3270 A>G位点GG基因型个体出生体长显著大于AA基因型，AG基因型个体出生胸围显著大于AA基因型（P<0.05）。【结论】 PLAG1基因在波尔山羊胎羊各组织中表达水平均高于成年母羊，发育早期的表达水平与出生羔羊体重相关。PLAG1基因可作为分子标记用于波尔山羊早期生长选育。

关键词: 波尔山羊; PLAG1基因; SNPs; 出生体重; 出生体尺

Abstract: 【Objective】 This study was aimed to investigate the sequence characteristics, polymorphisms of plemorphic adenoma gene 1 (PLAG1) gene and its effects on birth weight and body size in goats.【Method】 Boer goats were selected as the object, the PLAG1 gene were cloned and sequenced, the expression pattern of PLAG1 gene in various tissues of Boer goats at different ages and birth weight was identified by Real-time quantitative PCR, the PLAG1 protein level of leg muscle in lambs with different birth weight was detected by Western blotting.The SNPs in CDS and 3'-UTR of PLAG1 gene in Boer goats were further screened, and its correlation between different genotypes and birth weight and body size in Boer goats were analyzed.【Result】 The total length of PLAG1 gene CDS in Boer goats was 1 500 bp, encoding 499 amino acids.PLAG1 protein was relatively conservative.The tissue expression profile showed that the expression of PLAG1 gene in heart, small intestine and leg muscle in lambs with large birth weight were significant or extremely significantly higher than those with small birth weight, the expression of different tissues in fetal Boer goats was significantly or extremely significantly higher than that in adult ewes (P<0.05 or P<0.01). The expression of PLAG1 protein in leg muscles of lambs with large birth weight was extremely significantly higher than those with small birth weight (P<0.01).6 SNPs of PLAG1 gene 3'-UTR in Boer goats were screened, which were 2664 A>G, 2712 A>G, 2721 T>C, 2879 T>A, 2990 A>T and 3270 A>G, respectively.Correlation analysis showed that the birth tube circumference of AG genotype individuals at 2664 A>G was significantly longer than that of GG genotype (P<0.05);The birth length of TT genotype individuals at 2721 T>C was significantly longer than that of TC genotype (P<0.05);The birth tube circumference of TA genotype individuals at 2879 T>A was significantly longer than that of AA genotype (P<0.05);The birth weight of AA genotype individuals at 2990 A>T was significantly higher than that of AT genotype (P<0.05);The birth length of GG genotype individuals at 3270 A>G was significantly longer than that of AA genotype, and the birth chest circumference of AG genotype individuals at 3270 A>G was significantly larger than that of AA genotype (P<0.05).【Conclusion】 The expression of PLAG1 gene in different tissues of fetal goats were higher than those of adult ewes, the expression during early development was related to the birth weight.PLAG1 gene could be used as a molecular marker for early growth and breeding of Boer goats.

Key words: Boer goats; PLAG1 gene; SNPs; birth weight; body size

中图分类号:

S813.3

冯小品, 李隐侠, 张晨俭, 张俊, 孟春花, 钱勇, 曹少先. 波尔山羊PLAG1基因克隆、多态性鉴定及其与出生体重、体尺性状的关联分析[J]. 中国畜牧兽医, 2022, 49(9): 3465-3474.

FENG Xiaopin, LI Yinxia, ZHANG Chenjian, ZHANG Jun, MENG Chunhua, QIAN Yong, CAO Shaoxian. Coning and Polymorphism Identification of PLAG1 Gene and Its Association Analysis with Birth Weight and Body Size Traits in Boer Goats[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3465-3474.

参考文献

[1] KAS K, VOZ M L, RÖIJER E, et al.Promoter swapping between the genes for a novel zinc finger protein and[beta]|-catenin in pleiomorphic adenomas with t (3;8) (p21;q12) translocations[J].Nature Genetics, 1997, 15(2):170-174.
[2] DECLERCQ J, VAN DYCK F, VAN DAMME B, et al.Upregulation of IGF and Wnt signalling associated genes in pleomorphic adenomas of the salivary glands in PLAG1 transgenic mice[J].International Journal of Oncology, 2008, 32(5):1041-1047.
[3] VOZ M L, AGTEN N S, VAN DE VEN W J, et al.PLAG1, the main translocation target in pleomorphic adenoma of the salivary glands, is a positive regulator of IGF-Ⅱ[J].Cancer Research, 2000, 60(1):106-113.
[4] HENSEN K, BRAEM C, DECLERCQ J, et al.Targeted disruption of the murine PLAG1 proto-oncogene causes growth retardation and reduced fertility[J].Development, Growth & Differentiation, 2004, 46(5):459-470.
[5] ABI HABIB W, BRIOUDE F, EDOUARD T, et al.Genetic disruption of the oncogenic HMGA2-PLAG1-IGF2 pathway causes fetal growth restriction[J].Genetics in Medicine, 2018, 20(2):250-258.
[6] 郭潇潇, 李隐侠, 王悦, 等.湖羊PLAG1基因5'调控区多态性及其与早期体重的关联分析[J].畜牧兽医学报, 2021, 52(2):331-343. GUO X X, LI Y X, WANG Y, et al.Polymorphisms in the 5'regulatory region of PLAG1 gene and their association with early body weight of Hu sheep[J].Acta Veterinaria et Zootechnica Sinica, 2021, 52(2):331-343.(in Chinese)
[7] SONG Y X, XU L Y, CHEN Y, et al.Genome-wide association study reveals the PLAG1 gene for knuckle, biceps and shank weight in Simmental beef cattle[J].PLoS One, 2016, 11(12):e0168316.
[8] HOSHIBA H, SETOGUCHI K, WATANABE T, et al.Comparison of the effects explained by variations in the bovine PLAG1 and NCAPG genes on daily body weight gain, linear skeletal measurements and carcass traits in Japanese Black steers from a progeny testing program[J].Animal Science Journal, 2013, 84(7):529-534.
[9] LITTLEJOHN M, GRALA T, SANDERS K, et al.Genetic variation in PLAG1 associates with early life body weight and peripubertal weight and growth in Bos taurus[J]. Animal Genetics, 2011, 43(5):591-594.
[10] 李隐侠, 张俊, 钱勇, 等.湖羊NR5A1基因全序列克隆和表达特征研究[J].江苏农业学报, 2019, 35(1):114-121. LI Y X, ZHANG J, QIAN Y, et al.Full sequence cloning and expression characteristics of the ovine NR5A1 gene[J].Jiangsu Journal of Agricultural Sciences, 2019, 35(1):114-121.(in Chinese)
[11] PASCUAL-GARCIA A, ABIA D, MENDEZ R, et al.Quantifying the evolutionary divergence of protein structures:The role of function change and function conservation[J].Proteins, 2010, 78(1):181-196.
[12] SOUSOUNIS K, HANEY C E, CAO J, et al.Conservation of the three-dimensional structure in non-homologous or unrelated proteins[J].Human Genomics, 2012, 6(1):10.
[13] UTSUNOMIYA Y T, MILANESI M, UTSUNOMIYA A T H, et al.A PLAG1 mutation contributed to stature recovery in modern cattle[J].Scientific Reports, 2017, 7(1):17140.
[14] ZHOU Z H, HUANG B Z, LAI Z Y, et al.The distribution characteristics of a 19 bp indel of the PLAG1 gene in Chinese cattle[J].Animals (Basel), 2019, 9(12):1082.
[15] FINK T, TIPLADY K, LOPDELL T, et al.Functional confirmation of PLAG1 as the candidate causative gene underlying major pleiotropic effects on body weight and milk characteristics[J]. Scientific Reports, 2017, 7:44793.
[16] HOU J, QU K, JIA P, et al.A SNP in PLAG1 is associated with body height trait in Chinese cattle[J].Animal Genetics, 2020, 51(1):87-90.
[17] ZHONG J L, XU J W, WANG J, et al.A novel SNP of PLAG1 gene and its association with growth traits in Chinese cattle[J].Gene, 2018, 689:166-171.
[18] XU P, NI L, TAO Y, et al.Genome-wide association study for growth and fatness traits in Chinese Sujiang pigs[J].Animal Genetics, 2020, 51(2):314-318.
[19] QIAO R, GAO J, ZHANG Z, et al.Genome-wide association analyses reveal significant loci and strong candidate genes for growth and fatness traits in two pig populations[J].Genetics Selection Evolution, 2015, 47(1):17.
[20] RUBIN C J, MEGENS H J, MARTINEZ BARRIO A, et al.Strong signatures of selection in the domestic pig genome[J].Proceedings of the National Academy of Sciences of United States of America, 2012, 109(48):19529-19536.
[21] WEI Z Y, WANG K, WU H, et al.Detection of 15 bp deletion mutation within PLAG1 gene and its effect on growth traits in goats[J].Animals, 2021, 11(7):2064.