Biotechnologia Acta


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ISSN 2410-7751 (Print)
ISSN 2410-776X (Online)

Biotechnologia Acta
V. 8, No 5, 2015

"Biotechnologia Acta" V. 8, No 5, 2015
Р. 39-48, Bibliography 41, English
Universal Decimal Classification: 575.222.7:581.1


A.A.Peterson, M. Yu. Vasylenko, N. A. Matvieieva, M. V. Kuchuk

Institute of Cell Biology and Genetic Engineering of the National Academy of Sciences of Ukraine, Kyiv

Determination of the presence of the recombinant fusion protein (ESAT6-Ag85B(ΔTMD)-6His) and its accumulation level in duckweed plants (Lemna minor L.) was the aim of the research. ESAT6 and Ag85B are secretory proteins of Mycobacterium tuberculosis and are considered as potential candidates for development of new vaccine against tuberculosis (TB). Transgenic duckweed plants were obtained previously by Agrobacterium rhizogenes-mediated transformation and possessed fusion gene sequence esxA-fbpBΔTMD. Specific polyclonal antibodies were produced in immunized mice to identify levels of accumulation of TB antigens in plants. Recombinant antigen used for mice immunization was obtained in our laboratory by expression in E. coli. Western blot analysis revealed the recombinant tuberculosis antigen ESAT6-Ag85B(ΔTMD)-6His in extracts from transgenic L. minor plants. The level of accumulation of the protein corresponds to 0.4-0.5 µg protein per 1 g of fresh weight of plant. Additionally, the accumulation of recombinant protein was investigated in lyophilized transgenic plants after 1.5 year storage. Duckweed plants accumulating a recombinant analogue of M. tuberculosis secretory proteins can be used for development of plant-based edible vaccines.

Key words: Lemna minor, ESAT6, Ag85B(ΔTMD), genetic transformation, Agrobacterium rhizogenes.

© Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, 2008

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