双峰驼NGB基因克隆及序列分析

Cloning and Sequence Analysis of NGB Gene in Bactrian Camel

对双峰驼NGB基因进行克隆和序列分析,同时探讨双峰驼NGB的生理功能。在血样中提取双峰驼全基因组,采用分段TA克隆法获得NGB基因,运用多种软件对其基因序列及编码产物的结构、功能进行生物信息学分析。结果表明:双峰驼NGB基因序列全长5 210 bp,存在4个外显子(长度为89 bp、112 bp、120 bp、135 bp)、3个内含子(长度为1 505 bp、618 bp、1 437 bp)以及1个长度为257 bp的CpG岛。对双峰驼NGB肽链分析结果显示,包含所有基本氨基酸,各氨基酸数目相差较大,其中亮氨酸(L)占整个氨基酸组成的13.91%,天冬酰胺(N)占1.32%。双峰驼NGB相对分子质量为17 076.62,pI值5.39,半衰期30 h,不稳定系数56.50;蛋白整体呈现亲水性(-0.108),是可溶性蛋白。双峰驼NGB蛋白二级结构预测结果显示,α-螺旋所占比例76.16%,β-折叠所占比例0%,属于全α类蛋白质。双峰驼NGB在细胞骨架中占33.3%,在细胞质中占33.3%,在细胞核中占22.2%,在过氧化物酶体中占11.1%。双峰驼NGB与人亲缘关系最近(98.01%),与其他哺乳动物亲缘关系也较高(97.35%~94.03%),与斑马鱼亲缘关系较低(55.62%),说明NGB在哺乳动物长期进化过程中趋于保守。双峰驼NGB基因的成功克隆及序列分析,为进一步探究双峰驼应对极端环境过程中NGB基因可能发挥的生理功能提供理论基础。

To clone and sequence the NGB gene of Bactrian camel,and to explore the physiological function of Bactrian camel NGB.The NGB gene of Bactrian camel was obtained from the blood samples by segmented TA cloning method.The bioinformatics analysis of the structure and function of the gene sequence and coding products was carried out by using various software.The full-length NGB gene of Bactrian camel was 5 210 bp in length,with 4 exons(89 bp,112 bp,120 bp,135 bp in length)and 3 introns(1 505 bp,618 bp,1 437 bp in length)and 1 length 257 bp Cp G island. The analysis of the NGB peptide chain of Bactrian camel showed that all essential amino acids contained a large difference in the number of amino acids,among which leucine(L)accounted for 13.91% of the total amino acid composition,and asparagine(N)accounted for 1.32% of the total amino acid composition. The relative molecular mass of Bactrian camel NGB is 17 076.62,the p I value is 5.39,the half-life is 30 h,and the instability coefficient is 56.50.It is speculated that Bactrian camel NGB is unstable;the whole protein is hydrophilic(-0.108),which is soluble protein.The prediction results of NGB secondary structure showed that the proportion of α-helix was 76.16%,and the proportion of β-fold was 0%.It is lated lated that Bactrian camel NGB belongs to all α-protein.NGB accounts for 33.3% of the cytoskeleton,33.3% of the cytoplasm,22.2% of the nucleus,and 11.1% of the peroxisomes.Bactrian camel NGB has the closest relationship with humans(98.01%),and has a high genetic relationship with other mammals(97.35%-94.03%);it has a low genetic relationship with zebrafish(55.62%),indicating that the genetic relationship between species is more recently,the homology of NGB is higher. The successful cloning and sequence analysis of the Bactrian camel NGB gene provides a theoretical basis for further exploring the possible physiological function of the Bactrian camel during the response to extreme environments.