ANALYSIS OF STREPTOMYCIN-RESISTANCE OF ESCHERICHIA-COLI MUTANTS

被引:16
作者
BONNY, C [1 ]
MONTANDON, PE [1 ]
MARCMARTIN, S [1 ]
STUTZ, E [1 ]
机构
[1] UNIV NEUCHATEL,BIOCHIM VEGETALE LAB,CHANTEMERLE 18,CH-2000 NEUCHATEL,SWITZERLAND
关键词
RIBOSOME; S12; GENE; RIBOSOMAL-RNA; 16S; CHEMICAL FOOTPRINTING;
D O I
10.1016/0167-4781(91)90010-J
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We previously reported about Escherichia coli transformation experiments yielding streptomycin-resistant cells carrying a C912 to T transition in a plasmid-born 16S rRNA gene. These experiments were based on results obtained with streptomycin-resistant Euglena chloroplasts bearing an equivalent mutation in the single chloroplast 16S rRNA gene. We extended this study and transformed E. coli with plasmid constructs having a mutated 16S rRNA gene at position 914 (A to C) or a double mutation at positions 912 and 888 (C to T:G to A) or a mutation in the S12 gene (Lys-42 to Thr). We tested the transformed cells before and after a screening procedure in the presence of streptomycin. We find that the plasmid-born mutations protect colonies against a short streptomycin exposure, but ribosomes carrying mutated 16S rRNA do not significantly reduce codon misreading in vitro. However, ribosomes isolated from transformed cells after the screening procedure resist misreading. These ribosomes have acquired a second mutation in the S12 protein as shown in one case by sequencing and by transformation experiments. Furthermore, we show that the A914 to C mutation prevents (strongly reduces) base methylation in the central domain of 16S rRNA.
引用
收藏
页码:213 / 219
页数:7
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