Abstract

The Agrobacterium-mediated transformation of a duckweed plant (Lemna minor L.) with the use of the organogenic callus and nucleotide sequence of a gene for hirudin-1 optimized for expression in plants has been performed. Eight transgenic plant lines transformed by hirudin and seven lines transformed by the gene for β-glucuronidase were obtained. The expression of the glucuronidase gene was proven by histochemical staining and Western blot. ELISA of the transgenic plants showed that the content of β-glucuronidase in them varied from 0.28 to 1.43% of the total soluble protein. The expression of the hirudin-1 gene was confirmed by RT-PCR, with the maximum hirudin accumulation being equal to 0.02% of the total soluble protein. The results of the study can be used in the development of an expression system using the duckweed plant to obtain hirudin and other recombinant proteins for pharmaceutical use.

中文翻译：

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农杆菌介导的水Hi素和β-葡糖醛酸糖苷酶基因对小Lemna L.的转化

摘要

农杆菌介导的浮萍植物（小Lemna的转化）L.）已经利用器官发生的愈伤组织和针对植物中表达而优化的水-1素-1基因的核苷酸序列进行了研究。获得了由水rud素转化的8个转基因植物品系和由β-葡糖醛酸糖苷酶的基因转化的7个品系。葡糖醛酸糖苷酶基因的表达通过组织化学染色和Western印迹证实。转基因植物的酶联免疫吸附测定表明，它们中β-葡萄糖醛酸苷酶的含量为可溶性蛋白总量的0.28％至1.43％。通过RT-PCR证实了水rud素-1基因的表达，最大水hi素积累等于总可溶性蛋白的0.02％。该研究结果可用于开发使用浮萍植物的表达系统，以获得水rud素和其他重组蛋白，以用于制药。