杉木Cu/Zn-SOD基因克隆、序列特征及组织特异性表达  

Cloning,Sequence Characterization and Tissue Specific Expression of Cu/Zn-SOD Gene in Chinese Fir

作　　者：饶丽莎 许珊珊[1,2] 黄田盛 理挪 左丹丹[1,2] 曹光球[1,2] 林思祖[1,2] RAO Li-sha1,2 , XU Shan-shan1,2 , HUANG Tian-sheng1,2 , LI Nuo1,2, ZUO Dan-dan1,2, CAO Guang-qiuL2 ,LIN Si-zu1,2(1. College of Forestry, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China 2. State Forestry Administration Engineering Research Center of Chinese Fir ,Fuzhou, Fujian 350002, Chin)

机构地区：[1]福建农林大学林学院,福建福州350002 [2]国家林业局杉木工程技术研究中心,福建福州350002

出　　处：《西北林学院学报》2018年第2期75-82,共8页Journal of Northwest Forestry University

基　　金：国家林业局杉木工程技术研究中心平台建设(ptjh130002);中央财政支持地方专项(6213c0111);福建农林大学校重点项目建设专项"人工林培育学科建设"(6112c0310);林学院学科建设(6112c0300)

摘　　要：超氧化物歧化酶是植物体中的重要金属酶,Cu/Zn-SOD在超氧化物歧化酶中比例最大,它能够清除细胞内的超氧阴离子,保护机体免受氧化损伤。Cu/Zn-SOD基因对植物的抗氧化性有重要的作用,目前已证实在烟草、玉米等作物在逆境下抗氧化能力有所提高,但在杉木中的Cu/ZnSOD基因功能尚未深入研究。本研究以杉木组培苗为材料,通过RT-PCR技术,获得一段461bp的杉木Cu/Zn-SOD基因片段,运用生物信息学技术预测该基因蛋白的理化性质、亲疏水性、氨基酸组成、二三级结构、磷酸化结构及蛋白功能。结果表明,杉木Cu/Zn-SOD基因的ORF长度为312bp,编码104个氨基酸,预测蛋白理论等电点为5.61,分子量为15.287 91kD,是亲水性蛋白;同源氨基酸序列比对结果表明杉木Cu/Zn-SOD基因亚细胞定位预测该基因定位于细胞质内,Cu/Zn-SOD蛋白二三级结构无规则卷曲占取比例最大;预测未发现跨膜结构,在第18号氨基酸时为信号肽剪切位点,该蛋白属非分泌蛋白。杉木Cu/Zn-SOD基因的克隆对研究裸子植物及其分子机制具有重要意义,同时为植物Cu/Zn-SOD基因家族提供依据。Superoxide dismutase is an important metal enzyme in the plant, Cu/Zn-SOD was in the largest proportion of superoxide dismutase,it can remove the cell superoxide anion, protect the body to receive oxidative damage. Cu/Zn-SOD gene has an important effect on the antioxidant activity of plants. It has been confirmed that the antioxidant capacity of tobacco,maize and other crops is improved under adversity, but the function of Cu/Zn-SOD in fir is not deep. In this paper,Chinese fir tissue cultured seedlings were used as materials,Cu/Zn-SOD gene was cloned of 312 bp by RT-PCR method. The bio-informatics technique was used to predict the physicochemical properties, hydrophobicity, amino acid composition, structure, phosphorylation structure and protein function. The length of ORF of Cu/Zn-SOD gene was 312 bp and encoded 104 amino acids. The theoretical pI was 5.61 and the molecular weight was 15. 287 91 kD,which was a hydrophilic protein. The results of homologous amino acid sequence showed that Cu/Zn The subunit of SOD gene was predicted to be located in the cytoplasm,and the ratio of Cu/Zn-SOD protein was the largest in the case of random curl. It was predicted that the transmembrane structure was not found,and the signal peptide was cleaved at amino acid No. 18 site,which is a non-secreted protein.

关 键 词：杉木 Cu/Zn-SOD基因 基因克隆 生物信息学分析 

分 类 号：S791.27[农业科学—林木遗传育种]