ISSN 1995-5537
Р. 9-17, Bibliography 47, Russian.
Universal Decimal classification: 602.6:582.5/.9
Yu. S. Luchakivs’ka, O. M. Kishchenko, M. Yu. Vasylenko, Yu. V. Simonenko, N. V. Kuchuk
Institute of Cell Biology and Genetic Engineering of National Academy of Sciences of Ukraine, Kiyv
Recent studies have shown high immunogenicity of secretory Mycobacterium tuberculosis Ag85B and ESAT6 proteins. That's why it was suggested to use them as components of a new tuberculosis vaccine instead of the widely used BCG nowadays.
Production of recombinant proteins, especially vaccines, in transgenic plants has many economic and safety advantages as compared to animal and bacterial production systems, thus transgenic plants with raw-edible leaves, fruits and taproots could be discussed as so-called «edible vaccines». In this study we report the obtaining of carrot plants of Nantskaya, Karotel, Red Giant, Perfektziya and Amsterdamskaya varietes expressing the genes of secretory Mycobacterium tuberculosis proteins. The genes were transferred into plants via Agrobacterium-mediated transformation using Agrobacterium tumefaciens GV3101 and Agrobacterium rhizogenes A4 strains. The binary vectors contained genes of Ag85B, Ag85B(ΔTMD), ESAT6 and fused Ag85B(ΔTMD)-ESAT6 proteins driven by constitutive 35S CaMV promoter and root-specific Mll sugar beet promoter. PCR and RT-PCR analysis proved the presence and transcription for nptII selective gene and fbpB and esxA genes for all the analyzed carrot plants in case of nptII and for 92–95% of them in case of target genes. Western-blot analysis proved presence of the recombinant proteins for several plant lines.
Key words: carrot, recombinant protein, secretory proteins, Mycobacterium tuberculosis, Agrobacterium-mediated transformation.
© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2008