西藏牦牛ATGL基因克隆及生物信息学分析  

Cloning and bioinformatics analysis of the ATGL gene in Tibetan yak

作　　者：杨琴[1,2] 柴志欣[1,2] 宋乔乔[1,2] 张思源[1,2] 陈攀攀[1,2] 廖珂[1,2] 钟金城[1,2] 

机构地区：[1]动物遗传育种学国家民委一教育部重点实验室/西南民族大学,成都610041 [2]青藏高原研究院/西南民族大学,成都610041

出　　处：《黑龙江畜牧兽医》2015年第3期39-44,228,共7页Heilongjiang Animal Science And veterinary Medicine

基　　金：西南民族大学研究生创新型科研项目(CX2014SZ122)

摘　　要：为了研究西藏牦牛ATGL基因的理化特性及分子结构,试验采用PCR扩增和DNA测序技术以及生物信息学分析软件对西藏类乌齐牦牛ATGL基因进行克隆测序和生物信息学分析。结果表明:西藏类乌齐牦牛ATGL基因编码区全长为1 461 bp,可编码486个氨基酸,其中A、T、G、C碱基含量分别为17.2%、17.9%29.0%、35.9%,A+T为35.1%,G+C为64.9%;相对分子质量为53 417.9,理论等电点为6.83,在构成该蛋白所编码的氨基酸中亮氨酸(Leu)和脯氨酸(Pro)的含量最高,分别为13.4%和9.5%,总的带正电(Arg+Lys)和负电(Asp+Glu)残基分别为46和47;西藏类乌齐牦牛ATGL基因编码区核苷酸序列与普通牛、野猪、人、绵羊、山羊、绿头鸭、原鸡、小家鼠的核酸序列一致性分别为99.58%、87.52%、87.78%、96.26%、95.93%、67.63%、64.44%、82.65%;西藏类乌齐牦牛ATGL基因编码氨基酸序列与普通牛、野猪、人、绵羊、山羊、绿头鸭、原鸡、小家鼠的氨基酸序列一致性分别为99.79%、87.45%、87.66%、95.06%、96.09%、73.14%、69.56%、87.53%;ATGL蛋白为不稳定的跨膜蛋白,无信号肽;其二级结构主要以α-螺旋、β-折叠以及无规卷曲为主,其中94个氨基酸残基参与了16个α-螺旋的形成(占39.11%),35个氨基酸残基参与了9个β-折叠的形成;在三级结构中,其N端存在1个Patatin结构域。说明西藏牦牛ATGL基因在编码区序列存在明显的碱基偏倚性,在进化关系上ATGL基因无论核苷酸序列还是氨基酸序列都有较高的保守性。To study the physicochemical properties and molecular structure of the ATGL gene in Tibetan yak,the PCR amplification,DNA sequencing technology,and bioinformatics analysis software were used for the cloning,sequencing and bioinformatics analysis of the ATGL gene in Tibetan Leiwuqi yak. The results showed that the sequence of CDS of the ATGL gene was 1 461 bp in length,encoding for a protein of 486 amino acids; thereinto,the base contents of A,T,G,and C were 17. 2%,17. 9%,29. 0%,35. 9%,and the base contents of A ＋ T and G ＋ C were 35. 1% and 64. 9%,respectively; the relative molecular mass was 53 417. 9,and the theoretical isoelectric point was 6. 83. The contents of leucine（ Leu） and proline（ Pro） were the highest among the amino acids,which were 13. 4% and 9. 5%,respectively; the positively charged（ Arg ＋ Lys） residues and negative charged（ Asp ＋ Glu） residues were 46 and 47,respectively. The nucleic acid sequence of CDS of the ATGL gene of yak shared 99. 58%,87. 52%,87. 78%,96. 26%,95. 93%,67. 63%,64. 44%,and 82. 65% homology with that of Bos taurus,Sus scrofa,Homo sapiens,Ovis aries,Capra hircus,Anas platyrhynchos,Gallus gallus,and Mus musculus,respectively; the amino acid sequence of CDS of the ATGL gene of yak shared 99. 79%,87. 45%,87. 66%,95. 06%,96. 09%,73. 14%,69. 56%,87. 53%homology with that of Bos taurus,Sus scrofa,Homo sapiens,Ovis aries,Capra hircus,Anas platyrhynchos,Gallus gallus,and Mus musculus,respectively. The protein coded by yak ATGL gene,was unstable transmembrane protein,and had no signal peptide; the alpha- helix,beta-sheet and random curl mainly existed in its secondary structure,thereinto,94 amino acid residues involved in the formation of 16 alpha- helixes,which accounted for 39. 11%,and 35 amino acid residues involved in the formation of nine beta- sheets; in the tertiary structure,a Patatin structure domain existed in its n- terminal. The results indicate that there is an obvious base bias in the ATGL gene in the coding sequence,and the ATGL gene is more conse

关 键 词：西藏牦牛 ATGL基因 克隆测序 同源性比对 生物信息学分析 

分 类 号：S823.85[农业科学—畜牧学] Q785[农业科学—畜牧兽医]