[1]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.[doi:10.7685/jnau.201701006]
 LI Jie,ZHOU Hao,YU Jianfeng,et al.Molecular cloning and tissue expression analysis of VNN1 gene in goose[J].Journal of Nanjing Agricultural University,2018,41(1):140-146.[doi:10.7685/jnau.201701006]
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VNN1基因的克隆及组织表达分析()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
41卷
期数:
2018年1期
页码:
140-146
栏目:
出版日期:
2018-01-15

文章信息/Info

Title:
Molecular cloning and tissue expression analysis of VNN1 gene in goose
作者:
李洁12 周浩2 郁建锋2 姚文1 顾志良2
1. 南京农业大学动物科技学院, 江苏 南京 210095;
2. 常熟理工学院生物与食品工程学院, 江苏 常熟 215500
Author(s):
LI Jie12 ZHOU Hao2 YU Jianfeng2 YAO Wen1 GU Zhiliang2
1. College of Animal Sciences and Technology, Nanjing Agricultural University, Nanjing 210095, China;
2. College of Biological Science and Food Technology, Changshu Institute of Technology, Changshu 215500, China
关键词:
VNN1基因组织表达
Keywords:
Anser anserVNN1 genetissue expression
分类号:
S835.2
DOI:
10.7685/jnau.201701006
摘要:
[目的]Vanin是一种泛酰巯基乙胺酶,在脂类代谢中发挥重要作用。本试验目的是扩增鹅Vanin基因亚型(VNN1)基因的cDNA序列,检测VNN1基因在鹅不同组织中mRNA水平的表达规律。[方法]以太湖鹅(Anser anser)肝脏总RNA为模板,采用RT-PCR和快速扩增cDNA末端(RACE)方法克隆鹅VNN1基因全长cDNA序列,并运用多种生物信息学软件对其进行分析,应用实时荧光定量PCR(qPCR)技术检测VNN1基因在不同组织的表达情况。[结果]序列分析表明:鹅VNN1基因(GenBank登录号:KY399733)cDNA全长1 924 bp,包含1 476 bp的开放阅读框,35 bp的5’UTR和417 bp的3’UTR,共编码491个氨基酸。经预测鹅VNN1基因编码的蛋白质由7 646个原子组成,相对分子质量为54 870.61,理论等电点为5.23,是不稳定蛋白。在线预测发现鹅与鸭的VNN1蛋白三级结构呈现高度相似的螺旋和折叠。序列分析表明,鹅与绿头鸭的VNN1基因核苷酸及氨基酸同源性较高。系统进化树分析表明,鹅与绿头鸭亲缘关系较近。qPCR结果表明,鹅VNN1基因在肝脏的表达量显著高于小肠、肾、脾和肺(P<0.01)。[结论]获得鹅VNN1基因的全长cDNA序列,该基因在肝脏中高表达。
Abstract:
[Objectives]Vanin is a class of pantetheinase and plays an important role in lipid metabolism. The purpose of this study is to amplify the cDNA sequence of goose vanin subgenotype(VNN1) and detect the expression of VNN1 gene in different tissues. [Methods]In this study,the full-length cDNA of goose vanin subgenotype Vanin subgenotype (VNN1) gene was cloned by reverse transcription PCR and rapid amplification cDNA ends(RACE) with the total RNA from liver as template. The bioinformatics analysis and the expression detection of VNN1 mRNA in various tissues by real-time fluorescent quantitative PCR(qPCR) were performed,respectively. [Results]The results showed that the full-length cDNA of goose VNN1 gene(GenBank No.KY399733) was 1 924 bp in length,which contained 1 476 bp open reading frame,35 bp 5’UTR and 417 bp 3’UTR,encoding 491 amino acids. The VNN1 protein was predicted as a kind of unstable protein,which is composed of 7 646 atoms with the relative molecular weight of 54 870.61 and pI of 5.23. The three-dimensional structures of VNN1 protein prediction online between Anser anser and Anas platyrhynchos showed highly similar helix and fold. Sequence analysis showed that the nucleotide and amino acid homologies of VNN1 between Anser anser and Anas platyrhynchos were relatively high. Phylogenetic tree analysis showed the close affinity between Anser anser and Anas platyrhynchos. The qPCR results showed that the expression of goose VNN1 gene in the liver was significantly higher than that in the small intestine,kidney,spleen and lung(P<0.01). [Conclusions]This study has provided the full-length cDNA sequence of goose VNN1 gene,which is highly expressed in the liver.

参考文献/References:

[1] Duprè S,Graziani M T,Rosei M A,et al. The enzymatic breakdown of pantethine to pantothenic acid and cystamine[J]. Eur J Biochem,1970,16(3):571-578.
[2] Wittwer C T,Schweitzer C,Pearson J,et al. Enzymes for liberation of pantothenic acid in blood:use of plasma pantetheinase[J]. Am J Clin Nutr,1989,50(5):1072-1078.
[3] 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.
[4] Galland F,Malergue F,Bazin H,et al. Two human genes related to murine vanin-1 are located on the long arm of human chromosome 6[J]. Genomics,1998,53(2):203-213.
[5] 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.
[6] Caldwell R B,Kierzek A M,Arakawa H,et al. Full-length cDNAs from chicken bursal lymphocytes to facilitate gene function analysis[J]. Genome Biol,2005,6(1):R6.
[7] 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.
[8] Rommelaere S,Millet V,Gensollen T,et al. PPARalpha regulates the production of serum Vanin-1 by liver[J]. FEBS Lett,2013,587(22):3742-3748.
[9] van Diepen J A,Jansen P A,Ballak D B,et al. PPAR-alpha dependent regulation of vanin-1 mediates hepatic lipid metabolism[J]. J Hepatol,2014,61(2):366-372.
[10] Rakhshandehroo M. Peroxisome proliferator-activated receptor alpha target genes[EB/OL].[2010-8-9]. https://www.hindawi.com/journals/ppar/2010/612089.
[11] Chen S,Zhang W,Tang C,et al. Vanin-1 is a key activator for hepatic gluconeogenesis[J]. Diabetes,2014,63(6):2073-2085.
[12] Rommelaere S,Millet V, Manh T P V,et al. Sox17 regulates liver lipid metabolism and adaptation to fasting[J]. PLoS ONE,2014,9(8):e104925.
[13] 虞德兵,陆应林,徐昊翔,等. 基于线粒体COⅠ基因序列分析家鸭系统发育关系[J]. 南京农业大学学报,2011,34(6):109-114. DOI:10.7685/j.issn.1000-2030.2011.06.020. Yu D B,Lu Y L,Xu H X,et al. Analysis of phylogenetic relationship based on mitochondrial COⅠ gene sequence in domestic duck[J]. Journal of Nanjing Agricultural University,2011,34(6):109-114(in Chinese with English abstract).
[14] Maras B,Barra D,Duprè S,et al. Is pantetheinase the actual identity of mouse and human vanin-1 proteins[J]. FEBS Lett,1999,461(3):149-152.
[15] Li Y,Wang X,Yu J,et al. MiR-122 targets the vanin1 gene to regulate its expression in chicken[J]. Poult Sci,2016,95(5):1145-1150.
[16] 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 Lett,2000,483(2/3):149-154.
[17] Ruan B H,Cole D C,Wu P,et al. A fluorescent assay suitable for inhibitor screening and vanin tissue quantification[J]. Anal Biochem,2010,399(2):284-292.
[18] Motomura W,Yoshizaki T,Takahashi N,et al. Analysis of vanin-1 upregulation and lipid accumulation in hepatocytes in response to a high-fat diet and free fatty acids[J]. J Clin Biochem Nutr,2012,51(3):163-169.
[19] Leone T C,Weinheimer C J,Kelly D P. A critical role for the peroxisome proliferator-activated receptor alpha(PPARalpha) in the cellular fasting response:the PPARalpha-null mouse as a model of fatty acid oxidation disorders[J]. Proc Natl Acad Sci USA,1999,96(13):7473-7478.

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备注/Memo

备注/Memo:
收稿日期:2017-01-03。
基金项目:江苏省自然科学基金项目(BK20151257);苏州市科技支撑计划项目(SNG201245)
作者简介:李洁,硕士研究生。
通信作者:姚文,教授,博导,主要从事肠道微生物与动物营养和健康的研究,E-mail:yaowen67jp@njau.edu.cn;顾志良,教授,硕导,主要从事动物分子遗传学的研究,E-mail:zhilianggu88@hotmail.com。
更新日期/Last Update: 1900-01-01