人精氨酸酶Ⅰ基因克隆及生物信息学分析

Cloning and bioinformatics analysis of human-arginaseⅠ

[目的]克隆人精氨酸酶Ⅰ(human-ARGⅠ)基因并分析其生物学特性。[方法]根据NCBI数据库提供的人精氨酸酶Ⅰ基因序列设计其特异性引物,利用PCR扩增目的基因,并将其连接到pCMV-MYC载体上并利用生物信息学工具分析人精氨酸酶Ⅰ蛋白的生物学特性。[结果]酶切鉴定与DNA测序结果显示人精氨酸酶Ⅰ基因全长969bp,编码322个氨基酸残基;人精氨酸酶Ⅰ蛋白属于精氨酸酶———组蛋白去乙酰化酶超家族,是位于细胞质中的稳定亲水性蛋白,无信号肽,不含跨膜结构域,其二级结构由无规则卷曲,α-螺旋和延伸链构成,并且三级结构与二级结构预测结果高度一致,建模结果准确可靠。[结论]人精氨酸酶Ⅰ基因全长969bp,编码322个氨基酸残基,分子量为34734.94,pI6.72,其氨基酸序列中无信号肽,无跨膜结构域,是非分泌型及亲水性蛋白,是属于精氨酸酶-组蛋白去乙酰化酶超家族,无规则卷曲为二级结构中最主要的结构元件,并且三级结构与二级结构预测结果高度一致,建模结果准确可靠。

[Objective] To clone the human arginaseⅠ(human-ARGⅠ)gene and analyze its biological characters.[Methods]The specific primers were designed based on the human arginaseⅠ gene sequence from NCBI databases and target gene was amplified by PCR and ligated into the pCMV-MYC vector.The biological characters of human arginaseⅠ were further analyzed by using bioinformatics tools.[Results]The enzyme digestion and DNA sequencing results showed that the 969 bp length human arginaseⅠ gene was cloned which encoding the 322 amino acid residues.The human-ARGⅠ protein was belongs to arginase-a histone deacetylase superfamily,predicting a stable hydrophilic protein localized in the cytoplasm,which does not contain a signal peptide and transmembrane domains,which secondary structure was consists of random curl,α-helix and extended chain,and the predicting results of the tertiary structure and secondary structure were highly consistent,which show that the structure modeling result was reliable.[Conclusion]Human arginase Ⅰgene full-length was 969 bp,encoding 322 amino acids,molecular weight 34 734.94,pI 6.72,which does not contain the signal peptide and transmembrane domain,a hydrophilic non-secreted protein,which belongs to arginase-histone deacetylase superfamily,random coiling was the main structural element in its secondary structure,and the tertiary structure prediction result was highly consistent with that of secondary structure,which show that the structure modeling result was reliable.