论文部分内容阅读
以‘嘎拉’苹果为材料,克隆了6–磷酸葡萄糖酸脱氢酶基因Md6PGDH1(序列号:MDP0000279299)全长。序列分析显示,该基因包含一个长为1 047 bp完整的开放阅读框,编码347个氨基酸,分子量为36.430 k D,预测等电点为9.24。同源性分析表明Md6PGDH1还有另外3个同源基因;功能域分析表明Md6PGDH蛋白含有两个保守的绑定域;亚细胞预测表明Md6PGDH定位存在差异。分析Md6PGDH1启动子发现存在多个响应非生物胁迫的顺式作用元件。定量分析显示,Md6PGDH1在苹果的不同组织中都有表达,且受非生物胁迫诱导。原核诱导Md6PGDH1蛋白并进行蛋白酶活的测定,为后续蛋白功能鉴定奠定了基础。Md6PGDH1在苹果愈伤组织中过量表达,提高其抗盐胁迫的能力。
The full length of 6-phosphogluconate dehydrogenase gene Md6PGDH1 (SEQ ID NO: MDP0000279299) was cloned from ’Gala’ apple. Sequence analysis showed that the gene contained a complete open reading frame of 1 047 bp, encoding 347 amino acids with a molecular weight of 36.430 kD and a predicted isoelectric point of 9.24. Homology analysis showed that Md6PGDH1 has 3 other homologous genes. Functional domain analysis showed that Md6PGDH protein contained two conserved binding domains. Subcellular prediction indicated that Md6PGDH was located differently. Analysis of the Md6 PGDH1 promoter revealed the presence of multiple cis-acting elements in response to abiotic stresses. Quantitative analysis showed that Md6PGDH1 was expressed in different tissues of apple and induced by abiotic stress. Prokaryotic induction of Md6PGDH1 protein and determination of protease activity laid the foundation for the subsequent identification of protein function. Md6PGDH1 overexpression in apple callus increased its ability to resist salt stress.