Characterization of Temperature-sensitive Mutations Affecting the Ribosomal Proteins in Escherichia coli
| Project/Area Number |
59442002
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
遺伝学
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| Research Institution | Kobe University |
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Principal Investigator |
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| Project Period (FY) |
1984 – 1985
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| Project Status |
Completed (Fiscal Year 1985)
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| Budget Amount *help |
¥30,000,000 (Direct Cost: ¥30,000,000)
Fiscal Year 1985: ¥5,000,000 (Direct Cost: ¥5,000,000)
Fiscal Year 1984: ¥25,000,000 (Direct Cost: ¥25,000,000)
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| Keywords | E. coli K12 / Temperature-sensitive mutations / Ribosomal proteins / Gene cloning / <gamma^(delta)> -Insertion mutagenesis / Operon structure / リボゾーム蛋白質 |
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| Research Abstract |
The aim of this project was to characterize temperature-sensitive mutations in Escherichia coli that affected the structure and function of the ribosome. For this purpose a large number of temperature-sensitive mutants harbouring alterations in one or more of the ribosomal proteins have been isolated and genetically and biochemically characterized. It has thus been established that insertion of an IS1 element into the gene rpsE for ribosomal protein S5 as well as regional tandem multiplications in the gene rpsD for ribosomal protein S4 caused a temperature-sensitive phenotype, most probably by intefering with the continuation of translation of polycistronic messeges containing these ribosomal protein genes. Furthermore, the nucleotide sequence of a cluster of ribosomal protein genes including the genes for proteins S6, S18 and L9 has been determined. It was found that this cluster contained an open reading frame (ORF) capable of encoding a protein of 11.4 kd. Insertion mutagenesis of <gamma^(delta)> transposon revealed that this OrF did actually code for a protein with the expected molecular weight. In addition, the nucleotide sequence of another chromosomal region in which the gene rpmF for ribosomal protein L32 had been located was determined. The rpmF gene existed downstream of a gene encoding a 30 kd protein of unknown function with which the rpmF gene was concluded to form a transcriptional unit (operon). Furthermore, the nucleotide sequences of the genes coding for two acetylating enzymes, each specific for ribosomal protein S5 and S18, respectively, were determined. The two genes appeared to have no homology with each other. Furthermore, Southern hybridization analyses of them with the gene for a third acetylase specific for protein L12 showed that they had no homology with this gene, either. Thus, the work supported by the grant has established the genetic architecture of the ribosomal protein genes in E. coli.
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Research Products
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