论文部分内容阅读
PeaT1是从极细链格孢菌Alternaria tenuissima中分离的一种蛋白激发子,具有促进植物生长和诱导植物产生系统获得抗性的功能,为了实现peaT1基因在枯草芽胞杆菌Bacillus subtilis中的分泌表达,增加其应用途径,从枯草芽胞杆菌基因组DNA中分别扩增获得P43启动子和nprB基因的信号肽序列,并用SOE(Splicing by over lapping extension)方法与peaT1基因连接,将连接产物克隆到大肠杆菌-枯草芽胞杆菌穿梭表达载体pHY300-PLK上,构建了重组表达载体pHY43N-peaT1。将重组载体转化枯草芽胞杆菌WB800菌株,SDS-PAGE和Western blotting分析证实,在NprB信号肽的引导下,枯草芽胞杆菌成功分泌表达了PeaT1蛋白。构建的重组菌株能够显著增强幼苗抗旱性,提高小麦株高。
PeaT1 is a protein exciton isolated from Alternaria tenuissima. It has the function of promoting plant growth and inducing plant-derived system to acquire resistance. In order to achieve the secretory expression of peaT1 gene in Bacillus subtilis Bacillus subtilis, The signal peptide sequence of P43 promoter and nprB gene was amplified from Bacillus subtilis genomic DNA, and the signal peptide sequence of nprB gene was amplified by splicing by over lapping extension (SOE) method and the peaT1 gene was cloned into E.coli - Bacillus subtilis shuttle vector pHY300-PLK, a recombinant expression vector pHY43N-peaT1 was constructed. The recombinant vector was transformed into Bacillus subtilis WB800 strain, SDS-PAGE and Western blotting analysis confirmed that under the guidance of NprB signal peptide, Bacillus subtilis successfully secreted and expressed PeaT1 protein. The constructed recombinant strains can significantly enhance the seedling drought resistance and improve wheat plant height.