Comparison of translational enhancers in the course of in vitro synthesis of sheep pox viral coat proteins A27L and L1R

Abstract

In this work the abilities of plant viral and artificial enhancers of mRNA translation to increase synthesis of sheep pox viral coat proteins A27L and L1R in wheat germ cell-free system is studied and compared. For such a comparison 5' untranslated regions (UTRs) of genomic (g)RNAs of potato virus Y (PVY), tobacco etch virus (TEV), gRNA-4 of alfalfa mosaic virus (AMV), artificial enhancer 5xARC1, as well as 3' UTR of a tobacco mosaic virus (TMV) gRNA were used. The assessment of translation enhancement was made by measurement of activity of newly synthesized reporter protein β-glucuronidase (GUS) that hydrolyzes the fluorogenic substrate, as well as with the help of immunoblotting of proteins A27L and L1R. It is established that examined 5'-UTRs can considerably raise synthesis of proteins in comparison with arbitrary leader sequence (pl), which does not possess translational enhancer property.