斑马鱼神经介素S直系同源基因的克隆、序列分析及组织表达  

Cloning,Sequence Analysis and Tissue Distribution from Zebrafish(Danio rerio) Neuromedin S Precursor Gene

作　　者：陈文波[1] 张珍[1] 王琳凯[1] 

机构地区：[1]河南理工大学资源环境学院,焦作454000

出　　处：《生物技术通报》2012年第9期160-165,共6页Biotechnology Bulletin

基　　金：河南理工大学博士基金项目(B2010-7)

摘　　要：旨在更深入和广泛了解脊椎动物NMS前体基因的结构、进化和生理功能,利用生物信息学和分子生物学技术从斑马鱼脑组织中克隆得到了NMS前体基因序列,并对其基因和蛋白结构以及组织表达等方面进行了分析。结果表明,斑马鱼NMS前体基因cDNA序列编码110个氨基酸组成的多肽,其中包含22个氨基酸的信号肽。脊椎动物NMS前体蛋白序列比对结果显示,与哺乳动物不同,硬骨鱼类NMS前体蛋白在C端缺失很大一部分序列,不能形成类似哺乳动物的NMS成熟肽,经蛋白酶切割可能形成同源性较高的34个氨基酸的多肽。进化树和基因组分析显示,斑马鱼和青NMS前体蛋白聚类在一起,位于NMS这一分支上。同时,NMS前体基因在斑马鱼、青和人基因组中具有同线性关系。半定量RT-PCR结果显示,NMS前体基因在所检测的斑马鱼各组织中均有表达,其中以脑中表达量为最高。For further studying the structure, evolution and physiological funtions of vertebrate neuromedin S precursor gene, we cloned the cDNA sequence of neuromedin S precursor gene from the brain of zebrafish, and then analysed its gene and protein structures and tissue distribution. The results showed that the eDNA of NMS precursor gene had an open reading frame of 333 bp in length, encoding 110 amino acids which contained a signal peptide consisted of 22 amino acids. The alignment based on the amino acid sequences of vertebrate neuromedin S precursor proteins revealed that, unlike mammalian, the C-terminal of teleost neuromedin S precursor proteins had a deletion of some amino acids, and did not form the recognized mature NMS in mammal. A 34-residue neuropeptide could be produced from the NMS precursor protein by processing at the conserved first and second proteolytie processing sites. The phylogenetic tree analysis showed that zebrafish and medaka NMS precursor protein clustered firstly, and located the same group. Meanwhile, genomie analysis among zebrafish, medaka and human revealed that NMS precursor gene had the synteny relationship. By semi-quantitative RT-PCR, NMS precursor gene was expressed in all tissue examined, and the highest level was found in the brain.

关 键 词：斑马鱼 神经介素S 克隆 序列分析 表达 

分 类 号：Q78[生物学—分子生物学]