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Published on: January 18, 2014
[Effect of ribosomal protein mutation on the expression of alkaline protease gene in Bacillus subtilis]
Abstract:
Altogether 19 strains belong to 13 species of ribosomal protein mutants of Bacillus subtilis were tested in vitro transcription--translation system for their influence on the translation of alkaline protease gene (apr). It was found that 10 species (13 strains) of ribosomal protein mutants did affect the translation of apr mRNA. Streptomycin-dependent (Str-D) ribosomes almost did not translate the apr mRNA. Str-D inhibited the expression of apr gene at the translational level, but had no influence on neutral protease gene. There is a secondary structure complex at the translation initiation region of apr mRNA. When one of the secondary structure was destroyed by site directed mutagenesis, the translation efficiency was enhanced by 7.3 to 9.1 folds. The higher order structure of Str-D and Str-R ribosome were different and so were the affinity of Str-D and Str-R 30S subunits to the 5' end of apr mRNA. These results suggest that Str-D ribosomes could not translate apr mRNA because of the secondary structure complex, low initiation strength of apr mRNA, and alteration of the higher order structure of the Str-D ribosomes.
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