Journal of Bacteriology & Parasitology

Callus induction, regeneration and agrobacterium-mediated transformation of Wolffia arrhiza to obtain plant-producer of therapeutic proteins

9^th Biotechnology Congress

August 31-September 02, 2015 Orlando,Florida, USA

Khvatkov P1, Shvedova A1, Chernobrovkina M1, Pushin A1,2, Firsov A1,2, Shaloiko L2, Dolgov S1,2

1All-Russia Research Institute of Agricultural Biotechnology, Russia 2Institute of Bioorganic Chemistry, Russia

Posters-Accepted Abstracts: J Biotechnol Biomater

Abstract:

Vaccines creation based on transgenic plants may be considered as a groundbreaking technology in modern vaccinology with the advantages as compared to bacterial and yeast systems such as the lack of common human and animal pathogens and high level expression of heterologous proteins. To date the development of tissue culture systems in duckweeds is limited to species of the genus Lemna and Spirodella. Yet more promising target for biopharming is Wolffia arrhiza as an object for submerged cultivation in a fermenter. We have developed a two-step procedure of callus induction in Wolffia. At the first stage cluster structures are induced in the presence of 2,4-D and �?А during 16 weeks. At the second stage BA in the medium for callus induction is replaced by PCL over a period of 4 weeks. The created protocols for callus induction and regeneration allow achieving not only the high efficiency at each stage but also proceeding to the development of a protocol for Wolffia arrhiza stable transformation. The most efficient transgenesis and selection of the transgenic lines occur in the presence of hygromycin B. The successful transformation requires the presence both of 2,4-D and BA in the cultivation medium within 15 days. As a result of investigations 84 transgenic lines of Wolffia harboring both reporter [gfp (1 lines) and gus(3 lines)] and target [desulfatohirudin-1 (46 lines) and granulocyte colony-stimulating factor (34 lines)] genes were obtained. Integration of heterologous DNA was proved by molecular-biological analyzes (PCR and Southern blot analyzes). Expression of recombinant protein was confirmed by Western blot assay and ELISA.

Biography :

Email: khvatkov1987@gmail.com