ISSN 1995-5537
"Biotechnology" journal V. 4, No. 3, 2011
Р. 56-63, Bibliography 42, Ukrainian.
Universal Decimal classification: УДК 575.222.7:581.1
Inulin synthesis in chihory hairy roots
transformed with Agrobacterium rhizogenes
Matvieieva N. A., Kishchenko O. M., Shakhovsky A. M., Kuchuk M. V.
Institute of Cell Biology and Genetic Engineering of National Academy of Sciences of Ukraine, Kyiv
A fast reproducible transformation procedure employing Agrobacterium rhizogenes with genes of tuberculosis antigens ESAT6 and Ag85B from Mycobacterium tuberculosis or human interferon αб2b was developed for Cichorium intybus L. var foliosum Hegi. Obtained hairy roots have typical phenotype and were able to growth on hormone-free nutrient medium. Frequency of genetic transformation was 42.3% for construction рСВ161 with ifn-α2b gene and 5.9% when vector рСВ158 harbouring fused sequence of esxA::fbpBΔTMD genes coding for tuberculosis antigens was used. PCR and RT-PCR analysis have shown the presence and transcription of both selective nptII and target fbpBΔTMD, ifn-α2b genes in transformed hairy root lines. According to PCR data, all the analysed chicory root lines transferred TL-fragment Т-DNA plasmid pRi А4. Transgenic roots like untransformed ones accumulated inulin natural chicory, reserve polysaccharide, which synthesis is typical for chicory plants. The content of inulin in one of the obtained transgenic root lines was in 1.7 times more than in the control root line and was over 20% of dry weight.
.Key words: inulin, Cichorium intybus, Agrobacterium rhizogenes, genetic transformation, genes of tuberculosis antigenes ESAT6 and Ag85B or human interferon ?2b.
© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2008