Agrobacterium-Mediated Transformation of Lemna minor L. with Hirudin and β-Glucuronidase Genes

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.