普通烟草NtODB基因的电子克隆、表达及生物信息学分析

In silico cloning, expression and bioinformatics analysis of NtODB from common tobacco

摘要:

ODB基因在植物同源重组依赖性的DNA双键断裂修复过程中起重要作用，对植物诱变育种具有潜在的应用价值。克隆烟草NtODB基因并分析其表达特征为丰富ODB基因在同源重组DNA修复过程中的作用提供证据。为得到烟草NtODB基因序列，采用电子克隆技术获得该基因cDNA序列并克隆验证。进一步使用生物信息学方法分析该基因表达特征，对预测蛋白的理化性质、信号肽、高级结构等进行预测。生物信息学分析结果表明，NtODB基因开放阅读框包含579个碱基，蛋白含192个氨基酸残基，NtODB蛋白具有碱性和亲水性，主要定位于细胞质内；实时荧光定量PCR检测结果显示NtODB基因在不同组织中呈现组成型表达特征；亚细胞定位检测提示NtODB主要表达于细胞膜和叶绿体。NtODB基因的克隆与表达分析及其蛋白高级结构和理化性质的预测，可为进一步丰富ODB基因在同源重组依赖的DNA修复系统中的作用机制提供证据。

Abstract:

It has been reported that ODB genes play an important role in homologous recombination-directed DNA repair, suggesting their potential applications in plant breeding. To analyze the expression characteristics of tobacco NtODB gene, the cDNA sequence of NtODB was obtained using in silico cloning technique. The physicochemical properties, signal peptide, and advanced structures of the predicted protein were analyzed using bioinformatics tools. The results showed that the NtODB gene has a 579-bp open reading frame which encodes a protein with 192 amino acid residues. The protein NtODB is predicted to be alkaline and hydrophilic. Real-time quantitative PCR showed that NtODB was constitutively expressed in different tissues. Subcellular localization showed that NtODB was mainly expressed in cell membrane and chloroplast. These results may help us to better understand and elucidate the roles of ODB genes in the homologous recombination-directed DNA repair.