Cloning & Transgenesis
Open Access

Abstract

Genetically Transforming Russian Potato Cultivars for Resistance to Colorado Beetle

Anastasia M. Kamionskaya, Boris B. Kuznetsov, Vladimir Y. Ismailov, Vladimir D. Nadikta and Konstantin G. Skryabin

Using Agrobacterium-mediated transformation procedure, a total of 204 primary transformants of the Elizaveta, Lugovskoi and Nevski potato varieties carrying the cry IIIa gene were obtained. A number of biomolecular tests, including Southern hybridization, insert copy number screening, immunofluorescence analysis for assessment of the target gene expression levels, and PCR control for testing of the insert integrity, was carried out for these transformants, as well as the biosafety field trials for the transgenic lines selected during the biomolecular analysis. As a result, 3 transgenic lines (E2, N1, and L5) carrying one insert copy per genome with target gene expression level over 10 ppm were selected.