Abstract
The aim of research was to develop diagnostic system of BNYVV based on polymerase chain reaction for the identification of protein P75 of RNA-2. The bioinformative analysis of the nucleotide sequences of RNA-2 virus isolates of beet necrotic yellow vein (BNYVV) was made. The conserved sequences of a gene that encodes a protein 75 have been shown. Primers design for identification of RNA-2 has been made. It was synthesized pair primers: Forward 5’ – CTTTGGCAGGATTAGGCTCG-3’, Reverse 5’ – CACTCGGGACTATCACCAGG-3’ with amplification product fragments of size – 490 nucleotides pairs. The developed system was tested and confirmed the effectiveness of its use in Ukrainian model cDNA of BNYVV isolate. The annealing temperature of primer was optimized and we have found that a more optimal temperature for identification of BNYVV is in the range of 50–62°C. In further studies we plan to determine the nucleotide sequences of RNA-2 of Р75 protein and establish phylogenetic relationship to world′s isolates.
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