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Nucleic Acids Research, 1984, Vol. 12, No. 5 2243-2258
© 1984

MOLECULAR BIOLOGY

The effect of nucleotide analogs on cell-free gene expression

Michael Zimmer and Karl Heinz Scheit

Max-Planck-Institut für biophysikalische Chemie, Abteilung Molekulare Biologie Am Fassberg, 3400 Göttingen, FRG

Received January 13, 1984. Accepted February 15, 1984.

The effects of the followinq pyrimidine nucleoside 5'-triohosphates:f3 UTP, br5 UTP, r TTP, s2 UTP, s4 UTP and s2 CTP on cell-free expression of the ß-galactosidase gene in {lambda}h80dlac DNA as well as the galactokinase gene in plasmid 01–14 were investigated Only rTTP could substitute UTP in cell-free gene expression without restriction. Combinations of the other analogs with their respective natural congeners led to inhibition of gene expression. All analogs were found to inhibit transcription.Whereas br3 UTP and s4 UTP did not affect translation, RNA containing s2 UMP or s2 CMP residues respectively was found to function poorly in translation. Only in the case of f5 UTP could ambiguitive behaviour be demonstrated. Whether mispairing of f5 UMP residues, responsible for this ambiguity takes place in transcription or in translation, could not be decided.


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