Biopolymers and cell. 2007; 23 (6): 489 - 494

Construction of vectors for plant transformation to confer resistance to potyviruses

I. V. Korniichuk, V. P. Polischuk, S. D. Yeh

Genes HC-Pro, CIP, NIb, and CP are known to be the most conservative in genome of the potyviruses. The comparison of nucleotide sequences of these genes for 32 viruses of Potyvirus genus was carried out. The fragments with the highest percent of identity were selected, amplified from full size cDNA of Turnip mosaic virus (TuMV), and inserted into a high-copy TOPO-vector, used for the transformation of competent Escherichia coli cells. The construction of vectors for plant transformation on the basis of commercial vector pBI121 was performed. cDNA corresponding to one separate (HC-Pro, CIP, NIb or CP), two (HC-Pro and CIP), and three (HC-Pro, CIP, and NIb) conservative regions was inserted into the vector. The transformed cells of Agrobacterium tumefaciens carrying plasmids pBI121 were obtained.

Keywords: vector, cloned DNA (cDNA), Polymerase Chain Reaction (PCR), transformation, restriction analysis.