F3代波杂山羊VNN1基因多态性及其与产羔数的关联分析

doi:10.16431/j.cnki.1671-7236.2022.04.020

摘要/Abstract

摘要： 【目的】研究泛酰巯基乙胺酶-1(panthenoyl mercaptoethamine-1,VNN1)基因多态性与F3代波杂山羊产羔数的关系,从而从分子水平指导F3代波杂山羊的育种工作。【方法】选取107只F3代波杂山羊,采集血样提取DNA,根据VNN1基因(登录号:NC_030816.1)外显子序列设计7对引物,进行PCR扩增及测序,运用DNAStar软件分析测序结果,对可能存在的单核苷酸多态性(SNP)位点外显子全群进行PCR扩增测序,并进行Hardy-Weinberg平衡状态及遗传多样性分析;应用SPSS 16.0统计学软件对VNN1基因不同基因型与F3代波杂山羊产羔数进行关联分析。【结果】在VNN1基因在第5和7外显子中共发现7个SNPs位点:g.10956 A→G、g.11010 C→T、g.11103 C→T、g.11284 C→A、g.11313 T→C、g.18094 C→T和g.18158 G→C,且每个位点均存在3种不同基因型。χ²检验显示,g.10956 A→G、g.11010 C→T、g.11103 C→T、g.11313 T→C和g.18094 C→T位点均处于Hardy-Weinberg平衡状态,g.11284 C→A和g.18158 G→C位点均处于Hardy-Weinberg不平衡状态。多态信息含量均为中度多态((0.25<PIC<0.5)。关联分析发现,g.11010 C→T和g.11313 T→C位点与F3代波杂山羊产羔数显著相关,其中g.11010 C→T位点CC基因型产羔数显著低于CT和TT基因型,g.11313 T→C位点TT基因型产羔数显著低于TC和CC基因型(P<0.05);g.10956 A→G、g.11103 C→T和g.18158 G→C位点对F3代波杂山羊产羔数均无显著影响(P>0.05)。【结论】 VNN1基因在第5和7外显子中共发现7个SNPs位点,其中g.11010 C→T和g.11313 T→C位点对F3代波杂山羊产羔数有显著影响,VNN1基因可作为F3代波杂山羊的候选基因。

关键词: i>VNN1基因; F3代波杂山羊; 单核苷酸多态性(SNP); 产羔数; 关联分析

Abstract: 【Objective】 This experiment was conducted to investigate the relationship between panthenoyl mercaptoethamine-1 (VNN1) gene polymorphism and lambing number of F3 generation Boza goats,so as to provide molecular guidance for breeding of F3 generation Boza goats.【Method】 107 F3 generation Boza goats were selected for this experiment,blood samples were collected to extract DNA.According to the exon sequence of VNN1 gene(accession No.:NC_030816.1),7 pairs of primers were designed for PCR amplification and sequencing.DNAStar software was used to analyze the sequencing results,and PCR amplification and sequencing were performed on the entire group of possible single nucleotide polymorphism (SNP) sites.Hardy-weinberg equilibrium and genetic diversity analysis were carried out,and the correlation analysis between the different genotypes of VNN1 gene and the lamb number in F3 generation Boza goats was conducted by SPSS 16.0 statistical software.【Result】 There were 7 SNPs in exons 5 and 7 of VNN1 gene (g.10956 A→G,g.11010 C→T,g.11103 C→T,g.11284 C→A,g.11313 T→C,g.18094 C→T and g.18158 G→C),and 3 different genotypes existed in each site.χ² test showed that g.10956 A→G,g.11010 C→T,g.11103 C→T,g.11313 T→C and g.18094 C→T were in Hardy-Weinberg equilibrium.g.11284 C→A and g.18158 G→C were hardy-Weinberg disequilibrium.The polymorphic information content was moderate polymorphis (0.25<PIC<0.5).Correlation analysis showed that g.11010 C→T and g.11313 T→C were significantly correlation with the lamb number in F3 generation Boza goats,CC genotype of g.11010 C→T was lower than CT and TT genotypes;TT genotype of g.11313 T→C was significantly lower than TC and CC genotypes (P<0.05).There was no significant correlation between g.10956 A→G,g.11103 C→T and g.18158 G→C and the lamb number in F3 generation Boza goats (P>0.05).【Conclusion】 There were 7 SNPs in exons 5 and 7 of VNN1 gene,g.11010 C→T and g.11313 T→C of VNN1 gene were significantly correlated with the number of lambs in F3 generation Boza goats,and VNN1 gene could be used as a candidate gene.

Key words: VNN1 gene; F3 generation Boza goats; single nucleotide polymorphism (SNP); lambing number; correlation analysis

中图分类号:

S813.3

阮涌, 陈祥, 代林刚, 黄家锦, 安冬伟, 邹启顺. F3代波杂山羊VNN1基因多态性及其与产羔数的关联分析[J]. 中国畜牧兽医, 2022, 49(4): 1384-1392.

RUAN Yong, CHEN Xiang, DAI Lingang, HUANG Jiajin, AN Dongwei, ZOU Qishun. Correlation Analysis of VNN1 Gene Polymorphism and Lambing Number in F3 Generation Boza Goats[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(4): 1384-1392.

参考文献

[1] SILVESTRO D,GRAZIANI M T,ROSEI M A,et al.The enzymatic breakdown of pantethine to pantothenic acid and cystamine[J].European Journal of Biochemistry,2010,16(3):571-578.
[2] WITTWER C T,BURKHARD D,RIRIE K,et al.Purification and properties of a pantetheine-hydrolyzing enzyme from pig kidney[J].Journal of Biological Chemistry,1983,258(16):9733-9738.
[3] WITTWER C T,SCHWEITZER C,PEARSON J,et al.Enzymes for liberation of pantothenic acid in blood:Use of plasma pantetheinase[J].American Journal of Clinical Nutrition,1989,50(5):1072-1078.
[4] 李洁,周浩,郁建锋,等.鹅VNN1基因的克隆及组织表达分析[J].南京农业大学学报,2018,41(1):140-146. LI J,ZHOU H,YU J F,et al.Molecular cloning and tissue expression analysis of VNN1 gene in goose[J].Journal of Nanjing Agricultural University,2018,41(1):140-146.(in Chinese)
[5] 王中亮,李洁,姚文,等.泛酰巯基乙胺酶(Vanin-1)基因的研究进展[J].畜牧与兽医,2018,50(9):111-115. WANG Z L,LI J,YAO W,et al.Progress in research on the Vanin-1 gene[J].Animal Husbandry and Veterinary Medicine,2018,50(9):111-115.(in Chinese)
[6] AURRAND-LIONS M,GALLAND F,BAZIN H,et al.Vanin-1,a novel GPI-linked perivascular molecule involved in thymus homing[J].Immunity,1996,5(5):391-405.
[7] MARAS B,BARRA D,DUPRE S,et al.Is pantetheinase the actual identity of mouse and human Vanin-1 proteins?[J].FEBS Letters,1999,461(3):149-152.
[8] GRANJEAUD S,NAQUET P,GALLAND F.An ESTs description of the new Vanin gene family conserved from fly to human[J].Immunogenetics,1999,49(11-12):964-972.
[9] MARTIN F,MALERGUE F,PITARI G,et al.Vanin genes are clustered (human 6q22-24 and mouse 10A2B1) and encode isoforms of pantetheinase ectoenzymes[J].Immunogenetics,2001,53(4):296-306.
[10] CALDWELL R B,KIERZEK A M,ARAKAWA H,et al.Full-length cDNAs from chicken Bursal lymphocytes to facilitate gene function analysis[J].Genome Biology,2005,6(1):R6.
[11] ROMMELAERE S,MILLET V,GENSOLLEN T,et al.PPARalpha regulates the production of serum Vanin-1 by liver[J].FEBS Letters,2013,587(22):3742-3748.
[12] PITARI G,MALERGUE F,MARTIN F,et al.Pantetheinase activity of membrane-bound Vanin-1:Lack of free cysteamine in tissues of Vanin-1 deficient mice[J].FEBS Letters,2000,483(2-3):149-154.
[13] CHEN S,ZHANG W,TANG C,et al.Vanin-1 is a key activator for hepatic gluconeogenesis[J].Diabetes,2014,63(6):2073-2085.
[14] HOLLEBOOM A G,VERGEER M,HOVINGH G K,et al.The value of HDL genetics[J].Current Opinion in Lipidology,2008,19(4):385-394.
[15] GORING H H,CURRAN J E,JOHNSON M P,et al.Discovery of expression QTLs using large-scale transcriptional profiling in human lymphocytes[J].Nature Genetics,2007,39(10):1208-1216.
[16] JACOBO-ALBAVERA L,ROSA A,VILLARREAL-MOLINA T,et al.VNN1 gene expression levels and the G-137T polymorphism are associated with HDL-C levels in mexican prepubertal children[J].PLoS One,2012,7(11):e49818.
[17] KHAN M,DIAS F,SIRARD M A,et al.Transcriptome analysis of granulosa cells from growing dominant follicle reveals age-associated changes at the time of follicle selection in aged beef cattle[J].Reproduction,Fertility and Development,2012,25(1):148.
[18] BUNEL A,NIVET A L,BLONDIN P,et al.Cumulus cell gene expression associated with pre-ovulatory acquisition of developmental competence in bovine oocytes[J].Reproduction,2014,26(6):855-865.
[19] 郭应威,孙珂欣,田晨,等.分子标记技术在绵羊育种中的应用现状分析[J].中国畜牧兽医,2018,45(12):3505-3512. GUO Y W,SUN K X,TIAN C,et al.Application status analysis of molecular marker in sheep breeding[J].China Animal Husbandry & Veterinary Medicine,2018,45(12):3505-3512.(in Chinese)
[20] ALTSHULER D,POLLARA V J,COWLES C R,et al.An SNP map of the human genome generated by reduced representation shotgun sequencing[J].Nature,2000,407(6803):513-516.
[21] 曲娟娟,于敏,董蕾,等.SNP在动植物遗传育种中的应用[J].东北农业大学学报,2011,42(12):9-13. QU J J,YU M,DONG L,et al.Applications of SNP in animal and plant genetic breeding[J].Journal of Northeast Agricultural University,2011,42(12):9-13.(in Chinese)
[22] 朱向博,张丽.单核苷酸多态性及其在畜牧兽医领域的研究进展[J].现代畜牧兽医,2020,7:48-51. ZHU X B,ZHANG L.Review of single nucleotide polymorphism and research progress in animal husbandry[J].Modern Journal of Animal Husbandry and Veterinary Medicine,2020,7:48-51.(in Chinese)
[23] WANG J,SHETE S.Testing departure from Hardy-Weinberg proportions[J].Methods in Molecular Biology,2017,166(6):83-115.
[24] MAYO O. A century of Hardy-Weinberg equilibrium[J].Twin Research and Human Genetics,2008,11(3):249-256.
[25] 李碧春.动物遗传学[M].北京:中国农业大学出版社,2008. LI B C.Animal Genetics[M].Beijing:China Agricultural University Press,2008.(in Chinese)
[26] 陈浩林,杨红文,毛凤显,等.波尔山羊杂交改良贵州白山羊效果简介[A].第十七次全国动物遗传育种学术讨论会论文集[C].2013. CHEN H L,YANG H W,MAO F X,et al.Introduction to the effect of Boer goat hybrid improvement of Guizhou White goat[A].Proceedings of the 17th National Animal Genetics and Breeding Symposium[C].2013.(in Chinese)
[27] 焦仁刚,张明忠.波尔山羊、南江黄羊与贵州白山羊不同杂交组合生产性能比较[J].中国草食动物,2005,Z1:83-85. JIAO R G,ZHANG M Z.Comparison of production performance of Boer goat,Nanjiang Yellow goat and Guizhou White goat crosses[J].Chinese Journal of Herbivores,2005,Z1:83-85.(in Chinese)
[28] 杨光,宋正平,牟朝军.波尔山羊改良贵州沿河白山羊F1代羔羊肉用性能测定[J].中国草食动物,2003,23(3):21. YANG G,SONG Z P,MOU C J.Performance test of F1 generation lamb of Guizhou Yanhe White goat modified by Boer sheep[J].Chinese Journal of Herbivores,2003,23(3):21.(in Chinese)