小麦VP基因的克隆、生物信息学分析及功能初探

Cloning,Bioinformatics Analysis and Preliminary Functional Study of Wheat VP Gene

为研究小麦液泡膜H^+转运无机焦磷酸酶基因VP在植物中的作用,根据大麦、水稻等植物VP基因序列设计简并引物,用RACE法扩增小麦VP基因全长,对VP蛋白进行生物信息学分析,构建VP基因植物表达载体,转化拟南芥,对阳性转基因纯合体株系进行筛选和耐盐性鉴定。结果表明,小麦VP基因编码区序列全长2 286 bp,共编码761个氨基酸,等电点为5.17。VP蛋白为无信号肽的疏水性蛋白,主要含α螺旋。转VP基因拟南芥在NaCl胁迫下的萌发率、绿苗率均高于野生型拟南芥,成苗生长状态优于野生型拟南芥,说明小麦VP基因提高了转基因拟南芥的耐盐性。

In order to study the role of wheat vacuolar H +-pyrophosphatase gene VP in the plant,degenerate primers were designed according to the VP gene sequences of barley,rice and so on,and used to amplify the VP gene in wheat by RACE method.VP protein was analyzed by bioinformatics,and plant expression vector was constructed,then transfered into Arabidopsis thaliana .The positive transgenic homozygous lines overexpressing VP were screened,and their salt tolerance was identified.The results showed that full-length coding sequence of wheat VP gene was 2286 bp,encoded 761 amino acids,and the pI of VP protein was 5.17.VP protein is a hydrophobic protein without signal peptide,and mainly contains α helix.The germination rate and green seedlings rate of transgenic Arabidopsis thaliana overexpressing VP were higher compared with wild Arabidopsis thaliana ,and transgenic Arabidopsis thaliana overexpressing VP grew better compared with wild Arabidopsis thaliana under NaCl stress.These results showed that wheat VP gene increased tolerance to salt in transgenic Arabidopsis thaliana overexpressing VP .