摘要
为构建桃果胶乙酰酯酶(PpPAE)基因的原核表达载体pET-32a(+)-PpPAE,并获得稳定高效表达的重组蛋白,以青岛市现代农业质量与安全工程重点实验室前期获得的PpPAE基因(GenBank登录号KF017595)序列为依据,设计引物并扩增其cDNA片段,重组到原核表达载体pET-32a(+)中,将重组表达载体转化到大肠杆菌BL21(DE3)中,并用IPTG诱导表达,SDS-PAGE电泳分析表达产物。结果表明,已成功构建原核表达载体pET-32a(+)-PpPAE,重组质粒在0.1 mmol/L IPTG浓度下诱导4 h后,有大量融合蛋白表达,分子量约为64 kDa。为进一步研究该蛋白的功能奠定了基础。
This study was aimed to construct prokaryotic expression vector pET-32a（ ＋）-PpPAE and obtain high expression of PpPAE fusion protein in E. coli. cDNA sequence of PpPAE（ GenBank No. KF017595） was amplified and constructed into the prokaryotic expression vector pET-32a. Then the recombinant plasmid was transformed into E. coli BL21（ DE3） cell. The fusion protein was induced by IPTG and analyzed by SDS-PAGE. The result revealed that the prokaryotic expression vector of pET-32a（ ＋）-PpPAE was constructed successfully. The fusion protein of about 64 kDa was highly expressed and accumulated after induction with 0. 1 mmol / L IPTG for 4 h. This work laid the foundation for further study of Prunus persica PpPAE.
出处
《华北农学报》
CSCD
北大核心
2014年第2期18-21,共4页
Acta Agriculturae Boreali-Sinica
基金
国家青年基金项目(NSFC
30900976)
青岛农业大学第六届大学生创新教育立项项目
关键词
桃
果胶乙酰酯酶
原核表达
Prunus persica
Pectin acetylesterase（PAE）
Prokaryotic expression
