摘要
海藻糖-6-磷酸磷酸酶(TPP)是海藻糖合成的关键酶,通过合成具有渗透能力的海藻糖增加植物在逆境中的存活能力。本研究通过克隆得到水稻OsTPP3基因并构建原核表达载体进行体外表达,对基因启动子顺式作用元件组成、表达模式和模拟蛋白三维结构等生物信息学分析,揭示OsTPP3基因在水稻抵抗逆境胁迫中的作用。结果显示,OsTPP3基因编码366个氨基酸,预测分子量39.99 kD,成功构建原核表达载体并体外表达。OsTPP3蛋白具有三个保守的基序构成酶活性位点,蛋白两个结构域之间是底物结合位点。OsTPP3基因的基因芯片结果显示在正常生长条件下各器官均检测到表达信号,是组成型表达基因。但在花和种子各器官中表达量明显较高;7 d幼苗经干旱和盐胁迫处理后,OsTPP3基因在根和芽中表现出表达量明显增加的特点。OsTPP3基因启动子组成元件预测基因表达模式与基因芯片表达情况一致。通过对OsTPP3基因表达模式的研究,发现其在生长发育和渗透性胁迫中发挥重要功能,可作为转基因遗传育种的备选基因。
Trehalose-6-phosphatase（TPP） is the key enzyme in trehalose synthesis which can increase the survival ability of plants in adversity through synthesizing osmotic trehalose. OsTPP3 gene of rice was cloned and applied to construct prokaryotic expression vector expressed in vitro. The composition, expression pattern and three-dimensional structure of the simulated protein of cis-acting element of gene promoter were analyzed by bioinformatics to reveal the role of OsTPP3 gene in resistance to stress in rice. The results showed that OsTPP3 gene encoded 336 amino acid, with a predicted molecular mass of 39.99 k D, and successfully constructed prokaryotic expression vector expressed in vitro. OsTPP3 had three conserved motifs that const ituted the enzyme active site, and the substrate binding site was in the interface of two domains of OsTPP3 protein. Gene microarray data of OsTPP3 indicated that OsTPP3 was constitutive expression gene and all parts detected expression signal under normal growth condition, while OsTPP3 showed higher expression in each part of flower and seed. In addition, a marked increasing expression of OsTPP3 in seedling root and bud was observed in 7 d seedling after drought and salinity treatments. The expression pattern of the predicted gene in promoter components of OsTPP3 was consistent with the expression of gene microarray data. Through the analysis of the expression pattern of OsTPP3, we found that it played an important role in growth and osmotic stress, which could be used as a candidate gene for transgenic genetic breeding.
作者
杨雪
崔喜艳
李艳丽
杨美英
Yang Xue;Cui Xiyan;Li Yanli;Yang Meiying(College of Life Sciences, Engineering Research Center of the Chinese Ministry of Edueation for Bioreactor and Pharmaceutical Development, Jilin A- gricultural University, Changchun, 130118)
出处
《分子植物育种》
CAS
CSCD
北大核心
2018年第12期3785-3793,共9页
Molecular Plant Breeding
基金
吉林农业大学科研启动基金项目(201407)资助
