Test system for determination of HIV-1 frameshifting efficiency in animal cells.
We have developed a system in animal cells which allows the quantification of frameshifting determined by specific mRNA sequences. The method is based on the expression of an N-terminally extended firefly luciferase gene which requires frameshifting in order to be translated as a functional enzyme. The systems sensitivity is such that it allows the detection of even low efficiency of frameshifting. Our results show that the HIV-1 frameshift sequence including the 3' located stem-loop structure leads to ribosomal frameshifting at a lower level than that described for in vitro systems when tested in several fibroblastoid cell lines.
Reil
H
H
GBF-Gesellschaft für Biotechnologische Forschung, Braunschweig, F.R.G.
Hauser
H
H
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002872:Chromosome Deletion; D003062:Codon; D016368:Frameshift Mutation; D005822:Genetic Vectors; D015497:HIV-1 / Q000235:genetics*; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics*; D015183:Restriction Mapping; D012270:Ribosomes / Q000378:metabolism; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D003062:Codon; D012333:RNA, Messenger; D008156:Luciferases; D001616:beta-Galactosidase