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为了从抗逆性优良的牧草中挖掘新的耐盐、抗旱基因,进而为作物分子育种提供优良的基因资源,采用RACE(Rapid-Amplification of cDNA Ends)方法,克隆了披碱草的焦磷酸化酶基因EdHP1,其全长序列包含2 310 bp,编码770个氨基酸,含有14个跨膜区,是一个典型的膜蛋白,同时包含三个保守区(CS1、CS2和CS3)。氨基酸同源性比较发现,EdHP1与来自大麦等10种高等植物的焦磷酸化酶基因的同源性大于86%。EdHP1基因的表达受干旱、高盐和低温等非生物胁迫的诱导。功能分析证明,EdHP1基因能够显著提高转基因烟草对干旱、高盐胁迫的抗性,可用于作物的抗逆遗传改良。
In order to mine new salt-tolerant and drought-resistant genes from forage plants with excellent resistance, and to provide excellent genetic resources for crop molecular breeding, the pyrophosphorylation of Elymus sibiricus was cloned by RACE (Rapid-Amplification of cDNA Ends) The full-length EdHP1 gene contains 2 310 bp, encoding 770 amino acids and 14 transmembrane domains. It is a typical membrane protein and contains three conserved regions (CS1, CS2 and CS3). Amino acid homology comparison showed that EdHP1 shared more than 86% homology with pyrophosphorylase genes from 10 higher plants such as barley. The expression of EdHP1 gene is induced by abiotic stress such as drought, high salt and low temperature. Functional analysis showed that EdHP1 gene could significantly increase the resistance of transgenic tobacco plants to drought and salt stress and could be used to improve the resistance of crop plants.