类乌齐牦牛UCP3基因克隆与序列分析
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摘要
利用RT-PCR方法成功克隆UCP3基因,对其核苷酸序列及氨基酸序列进行生物学在线分析。结果表明,类乌齐牦牛UCP3基因CDS区序列长为936bp,A、G、C和T平均含量分别为20.83%、28.63%、31.09%和19.44%;与普通牛UCP3基因CDS区序列一致性高达99%;共发现碱基突变位点6个,其中同义突变4个,反义突变2个。UCP3基因共编码311个氨基酸,分子质量为34.18ku,理论等电位点9.53;其主要分布在线粒体内膜上,为非分泌蛋白;具有α-螺旋、无规则卷曲、延伸链、β-转角等结构。由系统进化树可知,类乌齐牦牛与野牦牛最接近,与普通牛次之。
The study was aimed to explore the characteristics of UCP3 in Leiwuqi yak, and its nucleotide and protein sequence were analyzed by online bioinformatics software.The CDS region of UCP3 gene was 936 bp, with the percentage of A, G, C and T was 20.83%, 28.63%, 31.09% and 19.44%, respectively;The similarity of the sequence is99%compared to cattle UCP3 gene;Six SNPs( 4 synonymous mutationand 2 missense mutation) were identified;and encoded 311 amino acids;the molecular weight andtheoretical equipotential point wwere34.18 ku and 9.53, respectively,mainly located in themitochondrial inner membranewithα-helix,Irregularcurl,extendedchain,andβ-turn;Leiwuqi yak UCP3 gene was closest associated with wild yak, was father with cattles. The full CDS region of the UCP3 gene of the Leiwuqi yak was successfully cloned, and it was conserved among different species.
The study was aimed to explore the characteristics of UCP3 in Leiwuqi yak, and its nucleotide and protein sequence were analyzed by online bioinformatics software.The CDS region of UCP3 gene was 936 bp, with the percentage of A, G, C and T was 20.83%, 28.63%, 31.09% and 19.44%, respectively;The similarity of the sequence is99%compared to cattle UCP3 gene;Six SNPs( 4 synonymous mutationand 2 missense mutation) were identified;and encoded 311 amino acids;the molecular weight andtheoretical equipotential point wwere34.18 ku and 9.53, respectively,mainly located in themitochondrial inner membranewithα-helix,Irregularcurl,extendedchain,andβ-turn;Leiwuqi yak UCP3 gene was closest associated with wild yak, was father with cattles. The full CDS region of the UCP3 gene of the Leiwuqi yak was successfully cloned, and it was conserved among different species.
引文
[1]蒋秋斐, 顾亚玲, 罗晓瑜,等. 肉牛生长速度功能基因研究进展[J]. 中国畜牧兽医, 2014, 41(7):192-197.
[2] Knoll A ,Putnová L, Dvorák J, et al. Linkage mapping of an AvaI PCR-RFLP within the porcine uncoupling protein 3 (UCP3) gene.[J]. Animal Genetics, 2010, 31(2):156-157.
[3] Stone R T, Smith T P. BovineUCP2 and UCP3 map to BTA15[J]. Animal Genetics, 2015, 30(5):378-381.
[4] Vidalpuig A, Solanes G, Grujic D, et al.UCP3: an uncoupling protein homologue expressed preferentially and abundantly in skeletal muscle and brown adipose tissue.[J]. BiochemBiophys Res Commun, 1997, 235(1):79-82.
[5] Sherman E L, Nkrumah J D, Murdoch B M, et al. Polymorphisms and haplotypes in the bovine neuropeptide Y, growth hormone receptor, ghrelin, insulin-like growth factor 2, and uncoupling proteins 2 and 3 genes and their associations with measures of growth, performance, feed efficiency, and carcass merit[J]. Journal of Animal Science, 2008, 86(1):1-16.
[6] Rousset S, Alvesguerra M C, Mozo J, et al. The Biology of Mitochondrial Uncoupling Proteins[J]. Diabetes, 2004, 53(Suppl 1):S130-5.
[7]Azzu V, Jastroch M, Divakaruni A S, et al. The regulation and turnover of mitochondrial uncoupling proteins[J]. BBA-Bioenergetics, 2010, 1797(6-7):785-791.
[8]Costford S R, Seifert E L, Bézaire V, et al. The energetic implications of uncoupling protein-3 in skeletal muscle.[J]. 2007, 32(5):884.
[9]Schrauwen P, Hesselink M.UCP2 and UCP3 in muscle controlling body metabolism.[J]. Journal of Experimental Biology, 2002, 205(15):2275.
[10]涂荣剑, 邓昌彦, 熊远著. 猪解耦联蛋白基因3(UCP3)3'侧翼区突变位点多态性及其对胴体、肉质性状的影响[J]. 畜牧兽医学报, 2004, 35(6):597-600.
[11]任亮, 朱宝芹, 韩丹,等. 猪UCP3基因多态性及其与脂肪沉积、肉质性状的相关分析[J]. 农业生物技术学报, 2006, 14(5):652-656.
[12]BartzM . Association studies on the porcine RETN,UCP1, UCP3 and ADRB3 genes polymorphism with fatness traits.[J]. Meat Science, 2009, 83(3):551-554.
[13]陈哲, 于峰祥, 于建宁, 等.猪UCP3基因第4内含子BccⅠ多态性检测及其与胴体和肉质性状的关联分析[J]. 江苏农业学报, 2012, 28(6):1338-1343.
[14]韩瑞华, 昝林森. 秦川牛及其杂种牛UCP3基因SNPs检测及其与胴体、肉质性状的相关性[J]. 畜牧兽医学报, 2009, 40(6):806-812.
[15]Zhang R , Li X . Association between IGF-IR, m-calpain andUCP-3 gene polymorphisms and growth traits in Nanyang cattle[J]. Molecular Biology Reports, 2011, 38(3):2179-2184.
[16] Wang Y, Yang W, Gui L, et al. Association and expression analyses of the UCP2and UCP3 gene polymorphisms with body measurement and meat quality traits in Qinchuan cattle[J]. Journal of Genetics, 2016, 95(4):1-8.
[17]蒋秋斐, 洪龙, 封元,等. 西门塔尔杂种牛UCP3基因多态性与生长性状的关联性分析[J]. 农业生物技术学报, 2018, 26(4):642-651.
[18]张娟, 母童, 蔡正云,等. UCP3基因多态性及与黑安格斯牛生长性状的关联分析[J]. 农业生物技术学报, 2018, 26(7):1195-1202.
[2] Knoll A ,Putnová L, Dvorák J, et al. Linkage mapping of an AvaI PCR-RFLP within the porcine uncoupling protein 3 (UCP3) gene.[J]. Animal Genetics, 2010, 31(2):156-157.
[3] Stone R T, Smith T P. BovineUCP2 and UCP3 map to BTA15[J]. Animal Genetics, 2015, 30(5):378-381.
[4] Vidalpuig A, Solanes G, Grujic D, et al.UCP3: an uncoupling protein homologue expressed preferentially and abundantly in skeletal muscle and brown adipose tissue.[J]. BiochemBiophys Res Commun, 1997, 235(1):79-82.
[5] Sherman E L, Nkrumah J D, Murdoch B M, et al. Polymorphisms and haplotypes in the bovine neuropeptide Y, growth hormone receptor, ghrelin, insulin-like growth factor 2, and uncoupling proteins 2 and 3 genes and their associations with measures of growth, performance, feed efficiency, and carcass merit[J]. Journal of Animal Science, 2008, 86(1):1-16.
[6] Rousset S, Alvesguerra M C, Mozo J, et al. The Biology of Mitochondrial Uncoupling Proteins[J]. Diabetes, 2004, 53(Suppl 1):S130-5.
[7]Azzu V, Jastroch M, Divakaruni A S, et al. The regulation and turnover of mitochondrial uncoupling proteins[J]. BBA-Bioenergetics, 2010, 1797(6-7):785-791.
[8]Costford S R, Seifert E L, Bézaire V, et al. The energetic implications of uncoupling protein-3 in skeletal muscle.[J]. 2007, 32(5):884.
[9]Schrauwen P, Hesselink M.UCP2 and UCP3 in muscle controlling body metabolism.[J]. Journal of Experimental Biology, 2002, 205(15):2275.
[10]涂荣剑, 邓昌彦, 熊远著. 猪解耦联蛋白基因3(UCP3)3'侧翼区突变位点多态性及其对胴体、肉质性状的影响[J]. 畜牧兽医学报, 2004, 35(6):597-600.
[11]任亮, 朱宝芹, 韩丹,等. 猪UCP3基因多态性及其与脂肪沉积、肉质性状的相关分析[J]. 农业生物技术学报, 2006, 14(5):652-656.
[12]BartzM . Association studies on the porcine RETN,UCP1, UCP3 and ADRB3 genes polymorphism with fatness traits.[J]. Meat Science, 2009, 83(3):551-554.
[13]陈哲, 于峰祥, 于建宁, 等.猪UCP3基因第4内含子BccⅠ多态性检测及其与胴体和肉质性状的关联分析[J]. 江苏农业学报, 2012, 28(6):1338-1343.
[14]韩瑞华, 昝林森. 秦川牛及其杂种牛UCP3基因SNPs检测及其与胴体、肉质性状的相关性[J]. 畜牧兽医学报, 2009, 40(6):806-812.
[15]Zhang R , Li X . Association between IGF-IR, m-calpain andUCP-3 gene polymorphisms and growth traits in Nanyang cattle[J]. Molecular Biology Reports, 2011, 38(3):2179-2184.
[16] Wang Y, Yang W, Gui L, et al. Association and expression analyses of the UCP2and UCP3 gene polymorphisms with body measurement and meat quality traits in Qinchuan cattle[J]. Journal of Genetics, 2016, 95(4):1-8.
[17]蒋秋斐, 洪龙, 封元,等. 西门塔尔杂种牛UCP3基因多态性与生长性状的关联性分析[J]. 农业生物技术学报, 2018, 26(4):642-651.
[18]张娟, 母童, 蔡正云,等. UCP3基因多态性及与黑安格斯牛生长性状的关联分析[J]. 农业生物技术学报, 2018, 26(7):1195-1202.