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23S Ribosomal RNA

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Heinz G. Floss – One of the best experts on this subject based on the ideXlab platform.

  • Overexpression of the Thiostrepton‐resistance Gene from Streptomyces azureus in Escherichia coli and Characterization of Recognition sites of the 23S rRNA A1067 2′‐methyltransferase in the Guanosine Triphosphatase Center of 23S Ribosomal RNA
    European journal of biochemistry, 1994
    Co-Authors: Andreas Bechthold, Heinz G. Floss

    Abstract:

    The thiostrepton-resistance gene encoding the 23S rRNA A1067 methyltransferase from Streptomyces azureus has been overexpressed in Escherichia coli using a T7-RNA-polymerase-dependent expression vector. The protein was efficiently expressed at levels up to 20% of total soluble protein and purified to near homogeneity. Kinetic parameters for S-adenosyl-l-methionine (Km= 0.1 mM) and an RNA fragment containing nucleotides 1029–1122 of the 23S Ribosomal RNA from E. coli (Km= 0.001 mM) were determined. S-Adenosyl-l-homocysteine showed competitive product inhibition (Ki= 0.013 mM). Binding of either thiostrepton or protein L11 inhibited methylation. RNA sequence variants of the RNA fragment with mutations in nucleotides 1051–1108 were tested as substrates for the methylase. The experimental data indicate that methylation is dependent on the secondary structure of the hairpin including nucleotide A1067 and the exact sequence U(1066)-A(1067)-G(1068)-A(1069)-A(1070) of the single strand.

  • overexpression of the thiostrepton resistance gene from streptomyces azureus in escherichia coli and characterization of recognition sites of the 23S rRNA a1067 2 methyltransferase in the guanosine triphosphatase center of 23S Ribosomal RNA
    FEBS Journal, 1994
    Co-Authors: Andreas Bechthold, Heinz G. Floss

    Abstract:

    The thiostrepton-resistance gene encoding the 23S rRNA A1067 methyltransferase from Streptomyces azureus has been overexpressed in Escherichia coli using a T7-RNA-polymerase-dependent expression vector. The protein was efficiently expressed at levels up to 20% of total soluble protein and purified to near homogeneity. Kinetic parameters for S-adenosyl-l-methionine (Km= 0.1 mM) and an RNA fragment containing nucleotides 1029–1122 of the 23S Ribosomal RNA from E. coli (Km= 0.001 mM) were determined. S-Adenosyl-l-homocysteine showed competitive product inhibition (Ki= 0.013 mM). Binding of either thiostrepton or protein L11 inhibited methylation. RNA sequence variants of the RNA fragment with mutations in nucleotides 1051–1108 were tested as substrates for the methylase. The experimental data indicate that methylation is dependent on the secondary structure of the hairpin including nucleotide A1067 and the exact sequence U(1066)-A(1067)-G(1068)-A(1069)-A(1070) of the single strand.

Andreas Bechthold – One of the best experts on this subject based on the ideXlab platform.

  • Overexpression of the Thiostrepton‐resistance Gene from Streptomyces azureus in Escherichia coli and Characterization of Recognition sites of the 23S rRNA A1067 2′‐methyltransferase in the Guanosine Triphosphatase Center of 23S Ribosomal RNA
    European journal of biochemistry, 1994
    Co-Authors: Andreas Bechthold, Heinz G. Floss

    Abstract:

    The thiostrepton-resistance gene encoding the 23S rRNA A1067 methyltransferase from Streptomyces azureus has been overexpressed in Escherichia coli using a T7-RNA-polymerase-dependent expression vector. The protein was efficiently expressed at levels up to 20% of total soluble protein and purified to near homogeneity. Kinetic parameters for S-adenosyl-l-methionine (Km= 0.1 mM) and an RNA fragment containing nucleotides 1029–1122 of the 23S Ribosomal RNA from E. coli (Km= 0.001 mM) were determined. S-Adenosyl-l-homocysteine showed competitive product inhibition (Ki= 0.013 mM). Binding of either thiostrepton or protein L11 inhibited methylation. RNA sequence variants of the RNA fragment with mutations in nucleotides 1051–1108 were tested as substrates for the methylase. The experimental data indicate that methylation is dependent on the secondary structure of the hairpin including nucleotide A1067 and the exact sequence U(1066)-A(1067)-G(1068)-A(1069)-A(1070) of the single strand.

  • overexpression of the thiostrepton resistance gene from streptomyces azureus in escherichia coli and characterization of recognition sites of the 23S rRNA a1067 2 methyltransferase in the guanosine triphosphatase center of 23S Ribosomal RNA
    FEBS Journal, 1994
    Co-Authors: Andreas Bechthold, Heinz G. Floss

    Abstract:

    The thiostrepton-resistance gene encoding the 23S rRNA A1067 methyltransferase from Streptomyces azureus has been overexpressed in Escherichia coli using a T7-RNA-polymerase-dependent expression vector. The protein was efficiently expressed at levels up to 20% of total soluble protein and purified to near homogeneity. Kinetic parameters for S-adenosyl-l-methionine (Km= 0.1 mM) and an RNA fragment containing nucleotides 1029–1122 of the 23S Ribosomal RNA from E. coli (Km= 0.001 mM) were determined. S-Adenosyl-l-homocysteine showed competitive product inhibition (Ki= 0.013 mM). Binding of either thiostrepton or protein L11 inhibited methylation. RNA sequence variants of the RNA fragment with mutations in nucleotides 1051–1108 were tested as substrates for the methylase. The experimental data indicate that methylation is dependent on the secondary structure of the hairpin including nucleotide A1067 and the exact sequence U(1066)-A(1067)-G(1068)-A(1069)-A(1070) of the single strand.

Suyeon Park – One of the best experts on this subject based on the ideXlab platform.

  • cost effectiveness of a tailored helicobacter pylori eradication strategy based on the presence of a 23S Ribosomal RNA point mutation that causes clarithromycin resistance in korean patients
    Journal of Gastroenterology and Hepatology, 2019
    Co-Authors: Jun-hyung Cho, Seong Ran Jeon, Hyun Gun Kim, So-young Jin, Suyeon Park

    Abstract:

    BACKGROUND AND AIM The Helicobacter pylori eradication rate using conventional triple therapy has decreased due to clarithromycin (CAM) resistance in H. pylori. Recently, dual priming oligonucleotide (DPO)-based multiplex polymerase chain reaction (PCR) can be used to detect H. pylori and point mutations in the 23S Ribosomal RNA gene causing CAM resistance. This study aimed to evaluate the success rate and cost-effectiveness of tailored H. pylori eradication using DPO-PCR. METHODS The H. pylori-positive patients diagnosed by a rapid urease test or DPO-PCR were enrolled from a single academic hospital. The patients with positive rapid urease test results received a CAM-based triple regimen. In the tailored therapy group that underwent DPO-PCR testing, patients with A2142G and/or A2143G point mutations were treated with a bismuth-containing quadruple regimen. The cost-effectiveness of H. pylori eradication success was evaluated according to the average cost per patient and the incremental cost-effectiveness ratio. RESULTS A total of 243 patients were allocated to the triple therapy group and 124 patients to the tailored therapy group. The first-line eradication rate of H. pylori was significantly higher in the tailored therapy group than in the conventional triple therapy group (92.7% vs 76.5%, P < 0.001). The average costs per patient for tailored therapy were $307.37 and $299.59 for first-line and second-line treatments, respectively. Compared with triple therapy, the incremental cost-effectiveness ratios of tailored therapy were $3.96 and -$3.81 per patient for first-line and second-line treatments, respectively. CONCLUSION In Korea, tailored H. pylori eradication using DPO-PCR may be more cost-effective than conventional triple therapy.

  • Cost-effectiveness of a tailored Helicobacter pylori eradication strategy based on the presence of a 23S Ribosomal RNA point mutation that causes clarithromycin resistance in Korean patients.
    Journal of gastroenterology and hepatology, 2018
    Co-Authors: Jun-hyung Cho, Seong Ran Jeon, Hyun Gun Kim, So-young Jin, Suyeon Park

    Abstract:

    BACKGROUND AND AIM The Helicobacter pylori eradication rate using conventional triple therapy has decreased due to clarithromycin (CAM) resistance in H. pylori. Recently, dual priming oligonucleotide (DPO)-based multiplex polymerase chain reaction (PCR) can be used to detect H. pylori and point mutations in the 23S Ribosomal RNA gene causing CAM resistance. This study aimed to evaluate the success rate and cost-effectiveness of tailored H. pylori eradication using DPO-PCR. METHODS The H. pylori-positive patients diagnosed by a rapid urease test or DPO-PCR were enrolled from a single academic hospital. The patients with positive rapid urease test results received a CAM-based triple regimen. In the tailored therapy group that underwent DPO-PCR testing, patients with A2142G and/or A2143G point mutations were treated with a bismuth-containing quadruple regimen. The cost-effectiveness of H. pylori eradication success was evaluated according to the average cost per patient and the incremental cost-effectiveness ratio. RESULTS A total of 243 patients were allocated to the triple therapy group and 124 patients to the tailored therapy group. The first-line eradication rate of H. pylori was significantly higher in the tailored therapy group than in the conventional triple therapy group (92.7% vs 76.5%, P