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℃、4 h,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 OsLEA19a ORF sequence,a pair of primers was designed,and the target 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 OsLEA19a peptide 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 OsLEA19a fusion protein exhibited improved resistance against diverse abiotic stresses：high salinity,hyperosmotic,heat and freezing.
出处
《华北农学报》
CSCD
北大核心
2012年第6期1-4,共4页
Acta Agriculturae Boreali-Sinica
基金
重庆市自然科学基金项目(cstc2012jjA80009)
重庆市教育委员会科学技术研究项目(KJ111108)
重庆高校创新团队建设计划(201040)
重庆三峡学院科学研究计划(11ZD-16)
