Kopertekh Lilya, Reichardt Sven
Institute for Biosafety in Plant Biotechnology, Julius Kühn-Institut (JKI) - Federal Research Centre for Cultivated Plants, Quedlinburg, Germany.
Front Plant Sci. 2021 Sep 10;12:712438. doi: 10.3389/fpls.2021.712438. eCollection 2021.
Transient expression in holds great potential for recombinant protein manufacturing due to its advantages in terms of speed and yield compared to stably transformed plants. To continue improving the quantity of recombinant proteins the plant host will need to be modified at both plant and cellular levels. In attempt to increase leaf mass fraction, we transformed with the gene, a positive regulator of the cell cycle. Phenotypic characterization of the T progeny plants revealed their accelerated above-ground biomass accumulation and enhanced rate of leaf initiation. In comparison to non-transgenic control the best performing line At-CycD2-15 provided 143 and 140% higher leaf and stem biomass fractions, respectively. The leaf area enlargement of the At-CycD2-15 genotype was associated with the increase of epidermal cell number compensated by slightly reduced cell size. The production capacity of the At-CycD2-15 transgenic line was superior to that of the non-transgenic . The accumulation of transiently expressed GFP and scFv-TM43-E10 proteins per unit biomass was increased by 138.5 and 156.7%, respectively, compared to the wild type. With these results we demonstrate the potential of cell cycle regulator gene to modulate both plant phenotype and intracellular environment of for enhanced recombinant protein yield.
与稳定转化的植物相比,瞬时表达在重组蛋白生产方面具有速度和产量优势,因而极具潜力。为持续提高重组蛋白的产量,需要在植物和细胞水平对植物宿主进行改良。为增加叶片质量分数,我们用细胞周期正向调节因子基因对植物进行转化。对T代植株的表型特征分析表明,它们地上生物量积累加速,叶片起始速率提高。与非转基因对照相比,表现最佳的株系At-CycD2-15的叶片和茎生物量分数分别高出143%和140%。At-CycD2-15基因型的叶面积增大与表皮细胞数量增加有关,细胞大小略有减小作为补偿。At-CycD2-15转基因株系的生产能力优于非转基因株系。与野生型相比,每单位生物量中瞬时表达的绿色荧光蛋白和单链抗体-TM43-E10蛋白的积累量分别增加了138.5%和156.7%。通过这些结果,我们证明了细胞周期调节基因在调节植物表型和植物细胞内环境以提高重组蛋白产量方面的潜力。