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Nucleic Acids Research, 1985, Vol. 13, No. 11 4113-4123
© 1985

Articles

Codon-defined ribosomal pausing in Escherichia coli detected by using the pyrE attenuator to probe the coupling between transcription and translation

Fons Bonekamp, Henrik Dalbtøge Andersen1, Thorkild Christensen1 and Kaj Frank Jensen*,

1Nordisk Gentofte, Niels Steensens vej 1 DK-2820 Gentofte, Denmark Institute of Biological Chemistry B, University of Copenhagen Sølvgade 83, DK-1307 Copenhagen K

*To whom correspondence should be addressed

Received April 18, 1985. Accepted May 9, 1985.

This communication describes an assay for the relative translation efficiency of individual codons which makes use of the pyrE attenuator to probe the coupling between transcription and translation at the end of an artificial leader peptide. By cloning of short synthetic DNA fragments the codons to be tested were placed in the middle of the leader peptide and the downstream transcription of a pyrE lacZ gene was monitored by measuring ß-galaetosidase activity. The substitution, one by one, of three AGG codons for arginine with three CGT codons for the same amino acid residue was found to cause a two fold increase per codon of transcription over the pyrE attenuator, such that an eight fold higher frequency of pyrE expression was seen when all three AGG codons were replaced by CGT codons. No such effect of codon composition was observed, when the cells were grown with a low UTP pool which causes a reduction of the mRNA chain growth rate


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