Biotechnologia Acta

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

"Biotechnologia Acta" V. 11, No 4, 2018
https://doi.org/10.15407/biotech11.04.073
Р. 73-83, Bibliography 25, English
Universal Decimal Classification: 604.6:577.218:575.113.3:582.926.2

THE EFFECT OF MONOCOT INTRONS ON TRANSGENE EXPRESSION IN Nicotiana GENUS PLANTS

I. O. Nitovska, M. Yu. Vasylenko, B. V. Morgun

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

The aim of the work was to study the effect of introns of the rice OsAct1 and the maize hsp70 genes on the transgene expression in Nicotiana plants in order to find out of their use in the testing of vectors containing these monocot introns. Next methods were used: Agrobacterium-mediated transformation of leaves of greenhouse N. benthamiana and N. tabacum plants by vector pCB271 containing the introns of cereals, light fluorescence microscopy and fluorimetry of GFP. The presence of transgenes was detected by polymerase chain reaction. The transient GFP expression was observed in infiltrated tissue of N. benthamiana. Transgenic plants of N. tabacum resistant to kanamycin were obtained. Fluorescence of GFP in extracts of some transgenic tobacco lines was shown. The impairment of the transgene expression in some N. tabacum transformants has been observed. So, transgenes, containing introns from the hsp70 corn or from the OsAct1 rice genes downstream the promotor, are expressed in Nicotiana plants. Thus, N. benthamiana and N. Tabacum plants can be used to test vectors constructs for cereals transformation. It has been shown that the monocot introns can have the negative impact on the transgene expression in Nicotiana plants.

Key words: . monocot introns, Nicotiana, Agrobacterium-mediated transformation, GFP.

© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2018

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