An in vivo dual-luciferase assay system for studying translational recoding in the yeast Saccharomyces cerevisiae.
A new in vivo assay system has been developed to study programmed frameshifting in the yeast Saccharomyces cerevisiae. Frameshift signals are inserted between the Renilla and firefly luciferase reporter genes contained in a yeast expression vector and the two activities are directly measured from cell lysates in one tube. Similar to other bicistronic reporter systems, this one allows the efficient estimation of recoding efficiency by comparison of the normalized activity ratios from each luciferase protein. The assay system has been applied to HIV-1 and L-A directed programmed -1 frameshifting and Ty1 and Ty3 directed +1 frameshifting. The assay system is amenable to high-throughput screening.
Harger
Jason W
JW
Department of Molecular Genetics, Microbiology and Immunology, UMDNJ Robert Wood Johnson Medical School, Piscataway, New Jersey 08854, USA.
Dinman
Jonathan D
JD
D000818:Animals; D001483:Base Sequence; D001517:Coleoptera; D018965:Frameshifting, Ribosomal; D017930:Genes, Reporter; D008156:Luciferases / Q000378:metabolism*; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D012333:RNA, Messenger; D008156:Luciferases