地黄超氧化物歧化酶(SOD)基因的克隆和生物学特性分析

Cloning and Characterization of Rehmannia glutinosa Superoxide Dismutase Gene

以地黄金九(03-2)叶片为材料,根据美国国家生物技术信息中心(National Center for Biotechnology Information,NCBI)中其他植物SOD基因设计简并引物,通过RT-PCR方法扩增出一个459 bp片段,该基因编码含152个氨基酸。采用生物信息学方法对该基因编码产物的二级结构、理化性质、亲水性/疏水性等方面进行预测分析。结果表明:克隆得到的基因为地黄SOD基因,该基因对应蛋白属于非跨膜的非分泌性亲水蛋白,二级结构具有54.61%的无规则卷曲,30.26%的延长链,9.21%的β-转角和5.92%的α-螺旋,与其他植物的SOD基因序列组成、蛋白性质结构等方面均具有高度相似性。该研究结果为进一步研究地黄SOD基因的表达和功能奠定了基础。

A 459 bp fragment encoding 152 amino acids is amplified by RT-PCR from the leaves of Dihuang Jinjiu(03-2)with degenerate primers designed according to the SOD gene of other plants in NCBI.The secondary structure,physical and chemical properties,hydrophilicity and hydrophobicity of the product encoded by the gene are predicted and analyzed by the bioinformatics method.The results shows that the cloned gene is Rehmannia glutinosa SOD,which is a non-transmembrane non-secretory hydrophilic protein.The secondary structure has 54.61%irregular curl,30.26% extension chain,9.21% β-rotation angle and 5.92% α-helix.It has high similarity with other plants in SOD gene sequence composition,protein structure and other aspects.These results lay a foundation for further study on the expression and function of SOD gene in Rehmannia glutinosa.