陈思涵, 钱靖, 彭杰军, 鲁宇文, 郑红英, 林林, 燕飞, 陈剑平. 农杆菌介导的外源基因在本氏烟中瞬时表达体系优化研究[J]. 西南林业大学学报, 2018, 38(4): 6-13. DOI: 10.11929/j.issn.2095-1914.2018.04.002
引用本文: 陈思涵, 钱靖, 彭杰军, 鲁宇文, 郑红英, 林林, 燕飞, 陈剑平. 农杆菌介导的外源基因在本氏烟中瞬时表达体系优化研究[J]. 西南林业大学学报, 2018, 38(4): 6-13. DOI: 10.11929/j.issn.2095-1914.2018.04.002
Sihan Chen, Jing Qian, Jiejun Peng, Yuwen Lu, Hongying Zheng, Lin Lin, Fei Yan, Jianping Chen. Optimization of Agroinfiltration-mediated Transient Gene Expression System in Nicotiana benthamiana[J]. Journal of Southwest Forestry University, 2018, 38(4): 6-13. DOI: 10.11929/j.issn.2095-1914.2018.04.002
Citation: Sihan Chen, Jing Qian, Jiejun Peng, Yuwen Lu, Hongying Zheng, Lin Lin, Fei Yan, Jianping Chen. Optimization of Agroinfiltration-mediated Transient Gene Expression System in Nicotiana benthamiana[J]. Journal of Southwest Forestry University, 2018, 38(4): 6-13. DOI: 10.11929/j.issn.2095-1914.2018.04.002

农杆菌介导的外源基因在本氏烟中瞬时表达体系优化研究

Optimization of Agroinfiltration-mediated Transient Gene Expression System in Nicotiana benthamiana

  • 摘要: 为提高外源蛋白的表达量及稳定性,期望通过RNA、蛋白水平的研究,制定一套高效的外源蛋白表达体系。通过共表达沉默抑制子、添加定位信号、沉默自噬关键基因atg5,分析各处理对植物外源蛋白表达的影响。结果表明:共表达植物病毒沉默抑制子p25或HC-Pro抑制mRNA降解速率,提高外源蛋白表达水平;外源蛋白增加内质网定位信号,可提高蛋白的持续表达能力;瞬时沉默自噬途径关键基因atg5,有利于外源蛋白的积累。外源蛋白表达过程中对易降解的外源蛋白,通过添加定位信号并结合使用沉默抑制子、自噬途径缺陷植株,可作为提高外源蛋白表达量及稳定性的新策略。

     

    Abstract: Our aim in this study is to obtain an effective plant expressing system for exogenous proteins by investigating the stability and accumulation of exogenous protein expressed in plant. Optimized exogenous proteins expressing system in plant is achieved via co-expression of silencing inhibitors, application of organelles localization signal and silencing of atg5. The results show that silencing suppressors p25 or HC-Pro of plant virus promoted exogenous protein accumulation by inhibiting mRNA silencing. Localization signal of endoplasmic reticulum (ER) enhanced exogenous protein yield. The silence of atg5, one of the key genes of autophagy pathway, increased the accumulation of exogenous protein by suppressing the degradation of exogenous protein. In view of the exogenous protein which is easy to be degraded, ER localization signal combined with silencing inhibitor or autophagy-deficient plants can be used as a new strategy to improve the expression and stability of exogenous proteins.

     

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