奶山羊超长链脂肪酸延伸酶6基因(ELOVL6)启动子的克隆及活性区域分析

Abstract
Figure/Table
References
Related Citation (15)

Download: PDF (2751 KB) HTML (1 KB)
Export: BibTeX | EndNote (RIS)

Abstract Abstract Elongase of very long chain fatty acids 6 (ELOVL6) is the rate-limiting enzyme in the elongation cycle in mammals that catalyzes the elongation of long-chain saturated and monounsaturated fatty acids such as 12–16 carbons. The objective of this study was to clone the active region of ELOVL6 promoter of Xinong Saanen Dairy Goat. Primers were designed based on genomic sequence of Yunnan black goat (Gene ID: NW_005100691.1 ), ELOVL6 promoter was cloned by PCR from goat genomic DNA. We used progressive deletion to get ten promoter fragments with different length , and then cloned into luciferase reporter gene vectors. The vectors and the pRL-TK were co-transfected into goat mammary epithelial cells. Using the dual-lueiferase reporter assay system to detect the expression activity．We obtained 2 370 bp（containing 2 168 bp upstream of the transcription start site）of 5' flanking sequence of ELOVL6 with 94%、81% and 80% homology with bovine, homo and mouse, respectively. Bioinformatics analysis showed that there was no typical transcription element TATA-box in ELOVL6 promoter. The core region of ELOVL6 promoter located in 109 to 40 bp upstream of the transcription start site and the potential negative control element was analyzed by deletion. Moreover, some consensus sites for PPAR, LXRA, SREBP-1, Sp1and NF-Y were identified in ELOVL6 promoter. Those result would lay a foundation for the study of function mechanism of ELOVL6 promoter.

Key words： Keywords Xinong Saanen dairy goat Elongase of very long chain fatty acids 6 (ELOVL6) promoter activity-analysis

Received: 17 November 2015 Published: 06 May 2016

Corresponding Authors: Jun LUO E-mail: luojun@nwsuaf.edu.cn

	Service

	E-mail this article
	Add to my bookshelf
	Add to citation manager
	E-mail Alert
	RSS
	Articles by authors
	SHI Qiu-Ya
	LUO Jun
	YAO Tai-Wei
	DIAO Bo
	YU Miao
	XU Hui-Fen
	YU Hui
	SHI Huai-Beng

Cite this article:

SHI Qiu-Ya,LUO Jun,YAO Tai-Wei, et al. Cloning and Characterization of Elongase of Very Long Chain Fatty Acids 6 (ELOVL6) Gene Promoter of Dairy Goat[J]. , 2016, 24(6): 790-798.

URL:

http://journal05.magtech.org.cn/Jwk_ny/EN/ OR http://journal05.magtech.org.cn/Jwk_ny/EN/Y2016/V24/I6/790

李君, 2011. 西农萨能羊脂肪酸合酶（FASN）基因启动子结构和功能研究(D).硕士学位论文,西北农林科技大学,导师：罗军,pp.18-23.（Li J.2011. Structural and functional analysis of fatty acid synthase gene promoter of xinong saanen dairy goat[D].Thesis for M.S.Northwest A&F University,Suppervisor：Luo J,pp.18-23.）
李君,葛婷,罗军,等. 2012. 山羊脂肪酸合酶基因(FASN)启动子结构与功能的初步分析[J].畜牧兽医学报,43(2):204-210.（Li J,Ge T,Luo J, et al. 2012. Structural and Functional Analysis of Fatty Acid Synthase Gene Promoter of Goat[J]. Acta Veterinaria et Zootechnica Sinica,43(2):204-210.）
萨姆布鲁克 J, 拉塞尔 D W. 2002. 分子克隆实验指南[M]. 科学出版社,北京.pp.463.( Sambrook J, Russell D W. 2002. The condensed protocols from moleculer cloning: a laboratory manual[M]. Science Press, Beijing.pp: 463. )
王莹, 韩烈保, 曾会明. 2007. 高等植物启动子克隆方法的研究进展[J]. 生物技术通报, 3: 97-100.(Wang Y,Han L B,Zeng H M. 2007. Advances on Studies of Plant Promoters[J].Biotechnology Bulletin,3:97-100.)
王桢，罗军，王伟，等. 2010. 奶山羊乳腺上皮细胞的分离、培养及鉴定[J]．生物工程学报, 26(8): 1123-1127.(Wang Z,Luo J,Wang W,et al. 2010. Characterization and culture of isolated primary dairy goat mammary gland epithelial cells [J].Chinese Journal of Biotechnology,26(8):1123-1127.)
吴敏, 罗军, 朱江江, 等. 2014 .奶山羊ELOVL6基因的克隆及其对4个脂肪酸代谢基因表达水平的影响[J]. 农业生物技术学报, 22(6): 672-680.(Wu M,Luo J,Zhu J J,et al.2014. Cloning of Elongase of Very Long Chain Fatty Acids 6 Gene(ELOVL6) and Its Effect on Four Genes Expression Involved in Fatty Acid Metabolism in Dairy Goat[J].Journal of Agricultugal Biotechnology,22(6):672-680.)
张晓，罗军，李建华，等. 2010. 西农萨能奶山羊脂肪酸合酶基因启动子的克隆及活性测定[J].中国农业科学，43(3):640-647.（Zhang X,Luo J,Li J H,et al.2010. Cloning and Activity Determination of Fatty Acid Synthase (FAS) Gene Promoter of Xinong Saanen Dairy Goat[J].Scientia Agricultura Sinica,43(3):640-647.）
Csak T, Ganz M, Pespisa J, et al. 2011. Fatty acid and endotoxin activate inflammasomes in mouse hepatocytes that release danger signals to stimulate immune cells[J]. Hepatology, 54(1): 133-144.
Corominas J, Ramayo-Caldas Y, Puig-Oliveras A, et al. 2013. Polymorphism in the ELOVL6 gene is associated with a major QTL effect on fatty acid composition in pigs[J]. PLoS ONE, 8(1): e53687.
Corominas J, Marchesi J AP, Puig-Oliveras A, et al. 2015. Epigenetic regulation of the ELOVL6 gene is associated with a major QTL effect on fatty acid composition in pigs[J]. Genetics Selection Evolution ,47:20.
Green C D, Olson L K.2011. Modulation of palmitate-induced endoplasmic reticulum stress and apoptosis in pancreatic -cells by stearoyl-CoA desaturase and Elovl6[J]. American Journal of Physiol Endocrinol Metabolism,300: E640–E649.
Iizuka K, Richard K, Bruick, et al. 2004. Deficiency of carbohydrate response element-binding protein (ChREBP) reduces lipogenesis as well as glycolysis[J]. PNAS, 101(19):7281-7286.
Jakobsson A, Westerberg R, Jacobsson A. 2006. Fatty acid elongases in mammals: their regulation and roles in metabolism[J]. Journal of Lipid Research, 45: 237-249.
Kumadaki S, Matsuzaka T, Kato T, et al. 2008. Mouse Elovl-6 promoter is an SREBP target[J]. Biochemical and Biophysical Research Communications, 368:261–266
Li J,Luo J , Xu H F, et al. 2015. Fatty acid synthase promoter: Characterization, and transcriptional regulation by sterol regulatory element binding protein-1 in goat mammary epithelial cells[J].Gene,561:157-164.
Matsuzaka T, Shimano H, Yahagi N, et al. 2002. Cloning and characterization of a mammalian fatty acyl-CoA elongase as a lipogenic enzyme regulated by SREBPs[J] . Journal of Lipid Research,43: 911-920.
Matsuzaka T, Shimano H. 2009.Elovl6: a new player in fatty acid metabolism and insulin sensitivity [J].J Mol Med,87:379–384
Mida K, Shamay A, Argov-Argaman N. 2012. Elongation and desaturation pathways in mammary gland epithelial cellsare associated with modulation of fat and membrane composition[J]. Journal of Agricultural and Food Chemistry, 60: 10657-10665.
Moon Y-A , Ochoa C R , Mitsche M A, et al. 2014. Deletion of ELOVL6 blocks the synthesis of oleic acid but does not prevent the development of fatty liver or insulin resistance. Journal of Lipid Research, 55: 2597–2605.
Park Y W, Ju′arez M, Ramos M,et al.2007. Physico-chemical characteristics of goat and sheep milk[J] . Small Ruminant Research , 68:88-113.
Saito R, Matsuzaka T, Karasawa T, et al.2011. Macrophage Elovl6 deficiency ameliorates foam cell formation and reduces atherosclerosis in low-density lipoprotein receptor-deficient mice[J]. Arteriosclerosis, Thrombosis, and Vascular Biology,31(9): 1973-1979.
Shimano H. 2012. Novel qualitative aspects of tissue fatty acids related to metabolic regulation:Lessons from Elovl6 knockout [J]. Progress in Lipid Research, 51: 267–271.
Sunaga H, Matsui H, Ueno M, et al. 2013. Deranged fatty acid composition causes pulmonaryfibrosis in Elovl6-deficient mice[J]. Nature communications, 4:2563
Sun H, Jiang T, Wang S, et al. 2013. The effect of LXRα, ChREBP and Elovl6 in liver and white adipose tissue on medium- and long-chain fatty acid diet-induced insulin resistance[J]. Diabetes Research and Clinical Practice, 102(3): 183-192.
Tang N, Matsuzaka T, Suzuki M, et al. 2014. Ablation of Elovl6 protects pancreatic islets from high-fat diet-induced impairment of insulin secretion[J]. Biochemical and Biophysical Research Communications,450:318-323.
Wang Y, Botolin D, Christian B, et al. 2005. Tissue-specific, nutritional, and developmental regulation of rat fatty acid elongases[J]. The Journal of Lipid Research, 46(4): 706-715.
Wang Y, Botolin D, Xu J, et al. 2006. Regulation of hepatic fatty acid elongase and desaturase expression in diabetes and obesity[J]. The Journal of Lipid Research, 47(9): 2028-2041.