Shyryna T. Small RNAs encoded in the genome regions of nuclear polyhedrosis viruses Bombyx mori and Autographa californica

The object: two sections of the genome DNA NPV Bmori and A. californica, containing the most recent (ph - orf1629) and complementary to them genes (p10 - p74). The aim: finding the possibility of participation of small RNAs in the regulation of gene expression A. californica NPV and B. mori, as well as to prove that these RNAs are essential structural component of polyhedra B. mori. The methods: bioinformatic methods to search for genes miRNAs in silico, biochemical methods for the isolation of RNA from polyhedra by agarose and polyacrylamide gels for analysis of RNA and RNA-protein complexes (RNP), polymerase chain reaction to identify the predicted miRNA precursors into RNA fractions of polyhedra, methods of cloning and sequencing of the amplified fragment to confirm it corresponds to the sequence of the predicted miRNA precursors. The results and novelty: the first time the algorithm search of genes by miRNAs secondary structure analysis of alternative transcripts. It was shown that the most recent genes A. californica NPV and B. mori encode miRNAs. It was shown that the gene NPV B. mori orf1629 encodes two small RNAs complementary to the mRNA orf ph polyhedrin. It was shown that B. mori NPV polyhedra include two small RNA nucleotide sequence and dimensions which match those of the small RNAs predicted by bioinformatics. It was shown that small RNA complexed with the N-terminal fragment of polyhedrin polypeptide chain. A hypothetical model of the high-RNP complex.