OsLEA2基因在原核生物中的异源表达和抗性分析

Analysis on Heterologous Expression of OsLEA2 Gene in Prokaryotes and Its Resistance

为了研究胚胎发育后期丰富蛋白基因OsLEA2的功能,利用PCR方法扩增目的基因,采用基因重组、蛋白质表达和抗逆性分析对OsLEA2基因进行研究。结果表明,构建的原核表达载体pET32a-OsLEA2,转化到E.coli BL21中,得到表达OsLEA2融合蛋白的工程菌;SDA-PAGE分析表明融合蛋白的分子量约为29.4 kDa。OsLEA2的表达增强了大肠杆菌对高盐、高温、低温和紫外照射等非生物胁迫的抗性。该研究为进一步改良作物品种、提高农作物的抗逆性奠定基础。

In order to investigate the function of late embryogenesis abundant protein gene OsLEA2, the target gene was amplified by PCR, and its OsLEA2 gene was studied with gene recombination technique, protein expression and analysis of stress tolerance. The result showed that the recombinant plasmid pET30a-OsLEA2 was constructed and transformed into E. coli BL21, and the genetic engineering bacteria that expressed OsLEA2 fusion protein were obtained. SDA-PAGE analysis indicated that the molecular mass of OsLEA2 fusion protein was 29.4 kDa. The results of stress tolerance assay demonstrated that recombinant E. coil cells producing OsLEA2 fusion protein exhibited improved resistance against diverse abiotic stresses： high salinity, heat, freeze-thaw and ultraviolet radiation. This research would provide a solid basis for the development of crops with enhanced resistance to diverse stresses.