We have measured the growth rates and elongation rates for different proteins in wild-type, miaA, rpsL, and miaA, rpsL double mutants of Escherichia coli in the presence as well as the absence of streptomycin. The data show that while miaA and rpsL mutants inhibit elongation rates to equivalent levels, miaA inhibits the growth rate twice as effectively as does rpsL. The double mutant is more effectively inhibited than either single mutant and Sm repairs in part the growth rate as well as protein elongation rates. The data suggest that the conditional streptomycin-dependent phenotype of the double mutant cannot be due simply to the depressed polypeptide elongation rates of the double mutant.

Petrullo et al. (1983) have studied the consequences of combining a mutation (rpsL-) that normally generates streptomycin resistant (Smr) ribosomes with a mutation (miaA-) that leads to loss of a tRNA hypermodification. They found surprisingly that such doubly mutant bacteria become streptomycin dependent (Smd). Here, we show in vitro that ribosomes purified from an Smr mutant behave very like Smd ribosomes when they are combined with tRNA from an miaA- mutant. Our analysis suggests that proofreading becomes excessively intense when the mutant components are combined, and that this reduces the efficiency of translation to the very low levels characteristic of Smd ribosomes. We show that Sm increases the efficiency of translation in vitro by suppressing the proofreading flows. We suggest that this will explain the growth stimulatory effect of Sm on the rpsL-, miaA- double mutants.

No abstract

No abstract

Methionine is one of the essential and first limiting amino acids in animal nutrition. In this study, an Escherichia coli methionine auxotroph bacterial strain that exhibits a linear growth response to methionine concentrations was transformed with a plasmid containing genes encoding ampicillin resistance and bioluminescence in order to develop a microbiological technique for methionine quantitation. Transformants were selected based on antibiotic resistance and plasmid containing candidates were confirmed by restriction enzyme digestion and gel electrophoresis. To confirm the bioluminescent phenotype, video imaging of the strain using long exposure photography yielded colonies exhibiting bioluminescence. The strain was also tested in the presence of ampicillin supplemented media with increasing methionine concentrations and growth response (measured as optical density, OD), growth rates and methionine affinities were compared before and after transformation. Although the transformed E. coli methionine auxotroph exhibited somewhat different growth kinetic responses than the nontransformed strain, the standard curves used for estimating methionine concentrations were not different. Based on the results in this study the transformed bioluminescent strain could be used as an OD‐based assay if bioluminescence equipment and materials are not available.