Fig. 7

Biological activity of recombinant RNase III enzyme and specificity assay. a Purified dsRNA molecules were incubated with the purified RNase III recombinant protein for 1, 2 and 4 min. Lane M: marker; Lanes 1–4: 0, 1, 2 and 4 min after incubation, respectively. b For the specificity assay, purified total RNAs by QIAZOL from the transformed E. coli BL21 with dsRNA coding sequence plasmid were incubated with the purified recombinant RNase III. To perform this test, 1 nmol of the purified dsRNA molecules were incubated for 1 min with ~5 ng of the recombinant purified RNase III enzyme in appropriate buffer for RNase III enzyme activity. Lane M: DNA ladder; Lane 1: non-incubated sample with the recombinant Asaia RNase III protein; Lanes 2 and 3: incubated RNAs with the recombinant Asaia RNase III protein. The incubated samples were loaded on agarose gel three folds higher than the non-incubated sample for better visualization and analyses of dsRNA cleavaging. These analyses revealed that the recombinant Asaia RNase III protein degrades the dsRNA molecules specifically and has no effect on ssRNA molecules. The produced dsRNA is marked in Lane 1 with a white arrow