牛BMPR Ⅰ A基因cDNA的克隆及组织表达谱分析

Cloning and tissue expression analysis of cDNA encoding the cattle BMPRⅠA gen

【目的】克隆牛BMPRⅠA基因cDNA全长,以进行生物信息学分析和组织表达谱研究。【方法】运用生物信息学方法,结合RT-PCR和SMART RACE技术,对牛BMPRⅠA基因进行cDNA全长克隆和生物信息学分析,并通过RT-PCR方法进行了组织表达谱研究。【结果】克隆得到了牛BMPRⅠA基因4 067 bp的cDNA全长序列,通过核酸序列分析发现,该基因编码532个氨基酸,与人、黑猩猩、小鼠、大鼠、犬和红原鸡在核酸序列上分别有91%,93%,89%,88%,91%和82%的同源性;在氨基酸序列上分别有97%,95%,96%,95%,97%和91%的同源性。通过生物信息学分析发现,牛BMPRⅠA蛋白包含一个信号肽序列和一个跨膜区序列,其成熟蛋白可能位于细胞膜上。在牛卵巢、肝、肌肉、小肠、脂肪、子宫、肾脏、心肌、肺、胰腺、睾丸、乳腺、瘤胃、脾脏、淋巴、胸腺等组织中都检测到有BMPRⅠA基因表达。【结论】牛BMPRⅠA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。

[Objective] The study was to clone and analyze the full-length cDNA sequence of the cattle BMPR Ⅰ A gene and the expression in 12 tissues. [Method] The BMPR Ⅰ A gene full-long eDNA was cloned by RT-PCR, and SMART RACE and analyzed by bioinformatics ways. [Result] The expressions were tested in 12 tissues by RT-PCR. The cattle BMPR Ⅰ A gene full-long eDNA is 4 067 bp,which encodes 532 amino acids and shares 91%,93% ,89%,88%,91% and 82% similarity in nucleic acid sequences and 97%,95%,96%,95%,97% and 91% similarity in amino acid sequences with human,chimpanzee, mouse,rat,dog and red jungle fowl,respectively. The cattle BMPR Ⅰ A protein has a signal peptide and a transmembrane domain. The mature protein is located in cell membrance by informatics analysis. The BMPR ⅠA gene are expressed in ovary, liver, skeleton muscle, small intestine, fat, uterus, kidney, heart, lung, pancreas,testis,galactophore,rumen,spleen,lymph and thymus gland. [Conclusion] The BMPR Ⅰ A is an important and conservative gene,which is broadly expressed in all kinds of tissues.