摘要摘要 超长链脂肪酸延伸酶6(elongase of very long chain fatty acids 6，ELOVL6)主要催化C12-C16饱和或单不饱和脂肪酸的延伸，是长链脂肪酸延长反应的限速酶。本研究根据云南黑山羊的基因组序列（ Gene ID:NW_005100691.1）设计引物，以血液DNA组织为模板，利用PCR技术克隆获得西农萨能奶山羊ELOVL6基因启动子的全长序列，通过生物信息学分析，构建了10个包含ELOVL6启动子不同缺失片段的荧光素酶报告基因载体，与pRL-TK内参载体共同转染乳腺上皮细胞，利用双荧光素酶系统检测不同片段的启动子活性。结果表明，克隆得到ELOVL6基因启动子全长序列2 370 bp(包含转录起始位点上游2 168bp)，与牛、人和鼠的基因组序列同源性分别为94%、81%和80%，ELOVL6启动子上无典型的真核生物启动转录元件TATA框。缺失突变研究发现，ELOVL6基因启动子前端存在负调控元件，启动子核心区域位于转录起始位点上游109～40 bp，该序列包含PPAR、LXR、SREBP-1、Sp1及NF-Y等转录因子的结合位点，为ELOVL6基因启动子功能研究奠定的基础。
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.
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