Khvatkov Pavel, Firsov Alexsey, Shvedova Anastasiya, Kozlov Oleg, Chernobrovkina Mariya, Pushin Alexander, Shaloiko Lyubov, Dolgov Sergey
All-Russia Research Institute of Agricultural Biotechnology, Moscow, Russia.
Nikita Botanical Gardens, Yalta, Russia.
3 Biotech. 2021 May;11(5):209. doi: 10.1007/s13205-021-02762-3. Epub 2021 Apr 10.
The production of recombinant proteins in transgenic plants is becoming an increasingly serious alternative to classical biopharming methods as knowledge about this process grows. , an aquatic plant unique in its anatomy, is a promising expression system that can grow in submerged culture in bioreactors. In our study 8550 explants were subjected to -mediated transformation, and 41 independent hygromycin-resistant lines were obtained, with the transformation efficiency of 0.48%. 40 of them contained the gene (codon-optimized for expression in plants) and were independent lines of nuclear-transformed , the transgenic insertion has been confirmed by PCR and Southern blot analysis. We have analyzed the accumulation of the target protein and its expression has been proven in three transgenic lines. The maximum accumulation of recombinant hirudin was 0.02% of the total soluble protein, which corresponds to 775.5 ± 111.9 ng g of fresh weight of the plant. The results will be used in research on the development of an expression system based on plants for the production of hirudin and other recombinant pharmaceutical proteins.
随着对转基因植物中重组蛋白生产过程的了解不断增加,这种生产方式正日益成为传统生物制药方法的一种重要替代方案。[此处缺失植物名称],一种在解剖结构上独特的水生植物,是一种有前景的表达系统,能够在生物反应器的浸没培养中生长。在我们的研究中,8550个外植体接受了[此处缺失介导方式]介导的转化,获得了41个独立的潮霉素抗性株系,转化效率为0.48%。其中40个株系含有[此处缺失基因名称]基因(针对植物表达进行了密码子优化),是核转化[此处缺失植物名称]的独立株系,转基因插入已通过PCR和Southern杂交分析得到证实。我们分析了目标蛋白的积累情况,并在三个转基因株系中证实了其表达。重组水蛭素的最大积累量为总可溶性蛋白的0.02%,相当于每克植物鲜重775.5±111.9纳克。这些结果将用于基于[此处缺失植物名称]植物开发用于生产水蛭素和其他重组药物蛋白的表达系统的研究。