牛BMP3基因cDNA克隆及组织表达谱分析

Cloning and tissues expression analysis of cDNA of the cattle BMP3 gene

用RT-PCR的方法克隆了鲁西黄牛BMP3基因1 555 bp的cDNA序列,该序列包含1个1 428 bp的完整开放阅读框,编码476个氨基酸残基;通过对推导的牛BMP3蛋白进行生物信息学分析发现,牛BMP3蛋白的分子量为53 506.27 D,等电点为9.42,该蛋白前22个氨基酸残基为信号肽序列,而亚细胞定位分析发现,该成熟蛋白可能位于细胞外.通过与最新公布的牛基因组比对,发现牛BMP3基因位于牛基因组第6号染色体上,由3个外显子和2个内含子组成,牛BMP3基因cDNA序列与人、小鼠、大鼠和鸡BMP3基因cDNA序列的相似性分别达到84%,81%,81%,79%,是进化上保守的基因.在牛卵巢、肝、肌肉、小肠、脂肪、子宫、肾脏、心肌、肺、胰腺等组织中都检测到BMP3基因表达,表明牛BMP3基因是一个功能重要、进化保守的基因,具有广泛的组织表达谱.

The Luxi cattle BMP3 gene cDNA was cloned by RT-PCR and a 1 555 bp cDNA sequence was obtained,including a 1 428 bp whole ORF region and encoding 476 animo acid residuals. The mo- lecular weight of predicted cattle BMP3 protein was 53 506.27 D and the isoelectric point was 9.42. The first 22 amino acids in N-terminal was signal peptide. The maturation protein was predicted to exist out of cells by subcellular localization analysis. The cattle BMP3 gene consists of three exons and two introns, which were located in the sixth chromatosome in contrast with the newest cattle genomes. Cattle BMP3 gene cDNA shared 84% ,81% ,81% ,79% similarity in nucleic acid sequences with human, rat, mouse and chicken. The BMP3 gene expression was detected in ovary, liver, skeleton muscle, small intestine, fat, uterus, kidney, heart, lung, pancreas. This indicated the cattle BMP3 gene is an important and conservative gene, which is broadly expressed in all kinds of tissues.