梅花鹿MRFAP1基因的编码区克隆及表达分析

Coding region cloning and expression analysis of MRFAP1 gene in Sika deer

为了探索梅花鹿MRFAP1基因的结构与生物学功能及其在梅花鹿鹿茸顶端间充质组织生长前中后期的表达差异,揭示MRFAP1对鹿茸生长重要的生物学意义,试验采用分子克隆技术成功获得了梅花鹿MRFAP1基因cDNA序列,根据梅花鹿MRFAP1基因cDNA序列,展开生物信息分析和梅花鹿鹿茸顶端间充质组织各生长阶段表达差异的研究。结果表明:获得的MRFAP1基因cDNA序列长532 bp,共编码127个氨基酸;梅花鹿MRFAP1蛋白是一种亲水不具跨膜结构的脂溶性不稳定蛋白分子,相对分子质量为14707.65;理论等电点(pI)为4.64;主要由α-螺旋组成,其次为无规卷曲。梅花鹿MRFAP1氨基酸序列与牛、东北虎、野猪相似性较高,而且MRFAP1基因在进化中高度保守;在梅花鹿鹿茸顶端间充质组织各生长阶段的表达量存在显著差异,其中在间充质组织生长前期表达量最高,中期表达量最低,前期的表达量是中期的(3.710±0.010)倍。说明成功克隆了梅花鹿MRFAP1基因,并分析得出MRFAP1基因在梅花鹿鹿茸顶端间充质组织各生长阶段的表达量规律为前期>中期>后期。

In order to explore the structure and biological function of MRFAP1 gene in Sika deer and the different expression level of MRFAP1 gene in the mesenchymal tissues of early,middle and last stages,the biological significance of MRFAP1 in the growth of pilose antler was revealed.This study successfully obtained the cDNA sequence of MRFAP1 gene by molecular cloning technology.According to the cDNA of MRFAP1 gene in Sika deer,the biological information analysis was carried out and the expression difference of MRFAP1 gene in the mesenchymal tissues of different stages was studied.The results showed that the cDNA sequence of MRFAP1 gene was 532 bp in length and encoded 127 amino acids in total.MRFAP1 protein was a hydrophilic lipid soluble unstable protein without transmembrane structure,and its relative molecular weight was 14707.65.The theoretical isoelectric point(pI)was 4.64.The prediction of the secondary structure of MRFAP1 protein was mainly composed of alpha-helix,followed by irregular curl.The amino acid sequence of MRFAP1 in Sika deer was highly similar to Bos taurus,Panther tigris altaica and Sus scrofa,and the MRFAP1 gene was highly conserved in evolution.The expression level of MRFAP1 gene was different in the mesenchymal tissues of different stages.The expression of MRFAP1 gene was the highest in the early stage,and the lowest in the middle stage,and the expression level in the early stage was(3.710±0.010)times as much as that in the middle stage.The above results indicated that the MRFAP1 gene was cloned successfully.And concluded that the expression pattern of MRFAP1 gene in the mesenchymal tissue of deer was early stage>middle stage>late stage.