ISSN 2410-776X (Online),
ISSN 2410-7751 (Print)
"Biotechnologia Acta" v. 6, no 6, 2013
https://doi.org/10.15407/biotech6.06.113
Р. 113-119, Bibliography 28, Ukrainian.
Universal Decimal classification: 581.143.6:58.085
GENETIC TRANSFORMATION AND ANALYSIS OF WHEAT TRANSGENIC CELL LINES BY IRAP-PCR
Bavol A. V.1, Dubrovna O. V.1, Morgun B. V.2
1Institute of Plant Physiology and Genetics of National Academy of Sciences of Ukraine, Kуiv
2Institute of Cell Biology and Genetic Engineering of National Academy of Sciences of Ukraine, Kуiv
The transgenic wheat cell lines were obtained via biolistic transformation of the callus cultures initiated from the 3-day-old sterile seedling shoot apexes. The pAHC25 vector construction used for 14- and 28-day-old callus cultures transformation carried the selective phosphinothricin-N-acetyltransferase (bar) gene and reporter ?-glucuronidase gene. The cell line selection was carried out on the media with phosphinothricin by means of graduated cell selection. The transgenic status of the obtained forms was proved by PCR-analysis. The presence of new relatively high molecular (more than 1 000 bp) amplicons were found out for three transformed lines by means of IRAP PCRanalysis with the primers coding for long termainal repeats sequences of SIRE 1 retrotransposon. This fact may prove transposition of this mobile genetic element. The new DNA fragments were detected for three of the seven analyzed lines but for the control callus. It is possible to assume at induction of SIRE 1 transposition bis probably caused by the genomic stress of foreign DNA inserting or associated with the transformation process (mechanical wounding, cultivation on selective media).
Key words: bread wheat, biolistic transformation, cell lines, the SIRE 1 retrotransposon, IRAP PCR.
© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2013
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