水牛FADS2基因的电子克隆及序列分析

doi:10.16431/j.cnki.1671-7236.2017.10.001

《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (10): 2829-2836.doi: 10.16431/j.cnki.1671-7236.2017.10.001

• 生物技术 • 下一篇

水牛FADS2基因的电子克隆及序列分析

马小娅, 庞春英, 梁莎莎, 陆杏蓉, 朱鹏, 段安琴, 梁贤威, 邓廷贤

中国农业科学院广西水牛研究所, 农业部(广西)水牛遗传繁育重点实验室, 南宁 530001

收稿日期:2017-04-05 出版日期:2017-10-20 发布日期:2017-10-20
通讯作者: 邓廷贤 E-mail:dtx282000@163.com
作者简介:马小娅(1989-),女,甘肃白银人,硕士,研究实习员,研究方向:水牛分子遗传学,E-mail:761195467@qq.com
基金资助:
广西科技重大专项（桂科AA16450002）；广西科技公关国合项目（桂科合1504001-3）；广西自然科学基金青年项目（2015GXNSFBA139103）；广西水产畜牧科技推广应用项目（桂渔牧科201633009）

Cloning and Bioinformatics Analysis of FADS2 Gene in Buffalo

MA Xiao-ya, PANG Chun-ying, LIANG Sha-sha, LU Xing-rong, ZHU Peng, DUAN An-qin, LIANG Xian-wei, DENG Ting-xian

Key Laboratory of Buffalo Genetics, Breeding Reproduction Technology, Ministry of Agriculture(Guangxi), Guangxi Buffalo Research Institute, Chinese Academy of Agricultural Sciences, Nanning 530001, China

Received:2017-04-05 Online:2017-10-20 Published:2017-10-20

摘要/Abstract

摘要：

试验旨在利用电子克隆法对水牛Δ6脂肪酸脱氢酶（Δ6-fatty acid desaturases，FADS2）基因进行克隆和生物信息学分析，为探究FADS2基因对水牛泌乳性能的作用机制奠定基础。以奶牛FADS2基因序列（GenBank登录号：NM_001083444.1）为探针设计引物，利用电子克隆法克隆水牛FADS2基因，并通过RT-PCR验证，对FADS2基因的序列特征进行生物信息学分析。测序结果表明，水牛FADS2基因序列全长为36 600 bp，由12个外显子和11个内含子组成，包含一个长1 335 bp的开放阅读框，可编码444个氨基酸。序列同源性分析显示，水牛FADS2基因编码序列与牦牛、黄牛、人、猪、家兔、虎鲸和褐家鼠序列的同源性分别为98.88%、98.88%、89.66%、90.79%、90.85%、92.35%和87.11%。蛋白质预测分析表明，水牛FADS2蛋白分子质量为52.51 ku，理论等电点（pI）为8.75，呈弱碱性，属于亲水性蛋白，无信号肽。系统进化树分析结果表明，FADS2基因在不同物种及进化的过程中具有高度保守性，其中水牛与牦牛、黄牛亲缘关系较近，与褐家鼠亲缘关系较远。水牛FADS2基因的成功克隆为今后阐明水牛泌乳性能的作用机制奠定了基础。

关键词: 水牛; FADS2基因; 电子克隆; 生物信息学分析

Abstract:

This study was aimed to clone the buffalo Δ6-fatty acid desaturases (FADS2) gene using in-Silico cloning and analyze its genetic struction with bioinformatics, which provide a foundation for investigating the milk performance in buffaloes. Primers were designed according the sequence of FADS2 gene in dairy cow (GenBank accession No.:NM_001083444.1). The FADS2 gene was amplified by RT-PCR, and its sequence was analyzed by bioinformatics. Sequence analysis revealed that the buffalo FADS2 gene had 36 600 bp in length and consisted of 12 exons and 11 introns, containing an open reading frame (ORF) of 1 335 bp which encoding 444 amino acids. Sequence homology analysis indicated that the buffalo FADS2 protein gene showed 98.88%, 98.88%, 89.66%, 90.79%, 90.85%, 92.35% and 87.11% identity with that of Bos mutus, Bos taurus, Homo sapiens, Sus scrofa, Oryctolagus cuniculus, Orcinus orca and Rattus norvegicus, respectively. Protein prediction analysis showed that the molecular weight and isoelectric point (pI) of buffalo FADS2 were 52.51 ku and 8.75, respectively, and the FADS2 protein was weak alkali and the hydrophobicity protein without signal peptide. Phylogenetic tree analysis showed that FADS2 gene was highly conserved in different species and evolutionary processes,buffalo was close to Bos mutus and Bos taurus, and was far from Rattus norvegicus. This study suggested that FADS2 gene was successfully cloned in buffalo, which laid a foundation for clarifying the mechanism of milk performance in buffalo.

Key words: buffalo; FADS2 gene; in-Silico cloning; bioinformatics analysis

中图分类号:

马小娅, 庞春英, 梁莎莎, 陆杏蓉, 朱鹏, 段安琴, 梁贤威, 邓廷贤. 水牛FADS2基因的电子克隆及序列分析[J]. 《中国畜牧兽医》, 2017, 44(10): 2829-2836.

MA Xiao-ya, PANG Chun-ying, LIANG Sha-sha, LU Xing-rong, ZHU Peng, DUAN An-qin, LIANG Xian-wei, DENG Ting-xian. Cloning and Bioinformatics Analysis of FADS2 Gene in Buffalo[J]. , 2017, 44(10): 2829-2836.

参考文献

[1] 胡皝,萧浪涛. 生物信息学在新基因全长cDNA电子克隆中的应用[J]. 生物技术通报,2007, 18(4):94-96.
[2] 孙淼,赵茂林. 利用表达序列标签电子克隆cDNA全序列的策略[J]. 生物技术通报,2010, 21(1):49-52.
[3] 卢冉. 鸡Δ6脂肪酸脱氢酶基因启动子区域多态性及基因时空表达的研究[D].郑州:河南农业大学,2011.
[4] 康相涛. 固始鸡-安卡鸡资源群构建及能量与脂肪代谢相关基因遗传变异的研究[D].兰州:甘肃农业大学,2009.
[5] Cho H P, Nakamura M T, Clarke S D. Cloning, expression, and nutritional regulation of the mammalian Delta-6 desaturase[J]. J Biol Chem, 1999, 274(1):471-477.
[6] Osman R H, Liu L, Xia L L, et al. Fads1 and 2 are promoted to meet instant need for long-chain polyunsaturated fatty acids in goose fatty liver[J]. Mol Cell Biochem, 2016, 418(7):103-117.
[7] 丁珍. 乳母乳汁多不饱和脂肪酸水平与FADS2基因多态性的关系研究[D].长春:吉林大学,2016.
[8] Ibeagha-Awemu1 E M, Akwanji K A, Beaudoin F, et al. Association between variants of FADS genes and omega-3 and omega-6 milk fatty acids of Canadian Holstein cows[J]. BMC Genetics, 2014, 15:25.
[9] 徐晨希,王梦琦,朱小瑞,等.中国荷斯坦牛FADS2基因3'端SNP突变对乳中脂肪酸组成的影响[J]. 中国农业科学,2016, 49(11):2194-2202.
[10] Nakayama K, Bayasgalan T, Tazoe F, et al. A single nucleotide polymorphism in the FADS1/FADS2 gene is associated with plasma lipid profiles in two genetically similar Asian ethnic groups with distinctive differences in lifestyle[J]. Human Genetics, 2010, 127:685-690.
[11] 邓廷贤,庞春英,陈明棠,等. 水牛信号转导子和转录激活子1基因电子克隆及序列分析[J]. 中国畜牧兽医,2014, 41(8):9-13.
[12] 刘仲娜,柴志欣,曾贤彬,等. 牦牛CAPN2基因的电子克隆及生物信息学分析[J]. 生物技术,2013, 23(1):42-45.
[13] Aki T, Shimada Y, Inagaki K, et al. Molecular cloning and functional characterization of rat delta-6 fatty acid desaturase[J]. Biochem Biophys Res Commun, 1999, 255(3):575-579.
[14] Marquardt A, Stohr H, White K, et al. Characterization cDNA cloning, genomic structure, and chromosomal localization of three members of the human fatty acid desaturase family[J]. Genomics, 2000, 66(2):175-183.
[15] Green C D, Ozguden-Akkoc C G, Wang Y, et al. Role of fatty acid elongases in determination of de novosynthesized monounsaturated fatty acid species[J]. J Lipid Res, 2010, 51:1871-1877.
[16] 朱士康. 鸡FADS2基因5'端SNP鉴定及启动子功能分析[D]. 郑州:河南农业大学,2014.
[17] Ashwell M S, Heyen D W, Sonstegard T S, et al. Detection of quantitative trait loci affecting milk production, health, and reproductive traits in Holstein cattle[J]. Journal of Dairy Science, 2004, 87(2):468-475.
[18] Alim M A, Wang P, Wu X P, et al. Effect of FASN gene on milk yield and milk composition in the Chinese Holstein dairy population[J]. Anim Genet, 2014, 45(1):111-113.
[19] Tanaka T, Shen J, Abecasis G R, et al. Genome-wide association study of plasma polyunsaturated fatty acids in the In CHIANTI study[J]. PLoS Genet, 2009, 5(1):e1000338.
[20] Nwankwo J O, Spector A A, Domann F E. A nucleotide insertion in the transcriptional regulatory region of FADS2 gives rise to human fatty acid delta-6-desaturase deficiency[J]. J Lipid Res, 2003, 44:2311-2319.
[21] Cobanoglu O, Zaitoun I, Chang Y M, et al. Effects of the signal transducer and activator of transcription 1(STAT1) gene on milk production traits in Holstein dairy cattle[J]. Journal of Dairy Science, 2006, 89(11):4433-4437.
[22] 洪雪莹. 鸡FADS2基因3'UTR区多态性检测及其与经济性状关联性分析[D]. 郑州:河南农业大学,2012.

水牛FADS2基因的电子克隆及序列分析

Cloning and Bioinformatics Analysis of FADS2 Gene in Buffalo

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