水稻OsLEA19a基因在大肠杆菌中的表达和抗逆性分析

摘要

根据胚胎发育后期丰富蛋白基因OsLEA19a的读码框序列设计引物,PCR扩增目的基因后,利用基因重组技术成功构建原核表达载体pET30a-OsLEA19a,并将重组质粒转化到E.coli BL21中.优化诱导OsLEA19a表达的条件,使目的蛋白可在E.coli BL21中高效表达.OsLEA19a融合蛋白的SDS-PAGE电泳分析表明,诱导蛋白表达的最适条件是37℃、4h,融合蛋白的大小大约为30 kDa.抗逆性分析表明,OsLEA19a的表达能增强大肠杆菌对高盐、高渗透压、高温和低温等非生物胁迫的抗性.%Based on late embryogenesis abundant protein gene 0sLEA19a ORF sequencer pair of primers was designed,and the targe gene was amplified by PCR. The recombinant expression plasmid pET30a-OsLEA19a was constructed and then transformed into E. coli BL21. Expression conditions of the plasmid encoded 0sLEA19a pep-tide in E. coli BL21 cells were optimized. The quantitative analysis of the fusion protein by SDA-PAGE showed that the optimal expression conditions were the expression temperature of 37℃ for 4 h. An about 30 kDa fusion protein was identified. The results of stress tolerance assay demonstrated that recombinant E. coil cells producing 0sLEA19a fusion protein exhibited improved resistance against diverse abiotic stresses ; high salinity, hyperosmotic, heat andrnfreezing.