The Experts below are selected from a list of 3264 Experts worldwide ranked by ideXlab platform

Robert M Goodman - One of the best experts on this subject based on the ideXlab platform.

  • Label free detection of 16s ribosomal rna hybridization on reusable dna arrays using surface plasmon resonance imaging
    Environmental Microbiology, 2002
    Co-Authors: Bryce P Nelson, Mark R Liles, Kendra B Frederick, Robert M Corn, Robert M Goodman
    Abstract:

    Summary In this paper, we describe the detection of bacterial cell-extracted 16S ribosomal RNA (rRNA) using an emerging technology, surface plasmon resonance (SPR) imaging of DNA arrays. Surface plasmon reso- nance enables detection of molecular interactions on surfaces in response to changes in the index of refraction, therefore eliminating the need for a fluo- rescent or Radioactive Label. A variation of the more common SPR techniques, SPR imaging enables detection from multiple probes in a reusable array format. The arrays developed here contain DNA probes (15-21 bases) designed to be complementary to 16S rRNA gene sequences of Escherichia coli and Bacillus subtilis as well as to a highly conserved sequence found in rRNAs from most members of the domain Bacteria. We report species-specific hybrid- ization of cell-extracted total RNA and in vitro tran- scribed 16S rRNA to oligonucleotide probes on SPR arrays. We tested multiple probe sequences for each species, and found that success or failure of hybrid- ization was dependent upon probe position in the 16S rRNA molecule. It was also determined that one of the probes intended to bind 16S rRNA also bound an unknown protein. The amount of binding to these probes was quantified with SPR imaging. A detection limit of 2 m g ml - - - 1 was determined for fragmented E.

  • Label free detection of 16s ribosomal rna hybridization on reusable dna arrays using surface plasmon resonance imaging
    Environmental Microbiology, 2002
    Co-Authors: Bryce P Nelson, Mark R Liles, Kendra B Frederick, Robert M Corn, Robert M Goodman
    Abstract:

    In this paper, we describe the detection of bacterial cell-extracted 16S ribosomal RNA (rRNA) using an emerging technology, surface plasmon resonance (SPR) imaging of DNA arrays. Surface plasmon resonance enables detection of molecular interactions on surfaces in response to changes in the index of refraction, therefore eliminating the need for a fluorescent or Radioactive Label. A variation of the more common SPR techniques, SPR imaging enables detection from multiple probes in a reusable array format. The arrays developed here contain DNA probes (15-21 bases) designed to be complementary to 16S rRNA gene sequences of Escherichia coli and Bacillus subtilis as well as to a highly conserved sequence found in rRNAs from most members of the domain Bacteria. We report species-specific hybridization of cell-extracted total RNA and in vitro transcribed 16S rRNA to oligonucleotide probes on SPR arrays. We tested multiple probe sequences for each species, and found that success or failure of hybridization was dependent upon probe position in the 16S rRNA molecule. It was also determined that one of the probes intended to bind 16S rRNA also bound an unknown protein. The amount of binding to these probes was quantified with SPR imaging. A detection limit of 2 micro g ml-1 was determined for fragmented E. coli total cellular RNA under the experimental conditions used. These results indicate the feasibility of using SPR imaging for 16S rRNA identification and encourage further development of this method for direct detection of other RNA molecules.

Gunnar Johansson - One of the best experts on this subject based on the ideXlab platform.

  • inhibition of the trichoderma reesei cellulases by cellobiose is strongly dependent on the nature of the substrate
    Biotechnology and Bioengineering, 2004
    Co-Authors: Marju Gruno, Priit Valjamae, Goran Pettersson, Gunnar Johansson
    Abstract:

    The inhibition effect of cellobiose on the initial stage of hydrolysis when cellobiohydrolase Cel 7A and endoglucanases Cel 7B, Cel 5A, and Cel 12A from Trichoderma reesei were acting on bacterial cellulose and amorphous cellulose that were [(3)H]- Labeled at the reducing end was quantified. The apparent competitive inhibition constant (K(i)) for Cel 7A on [(3)H]-bacterial cellulose was found to be 1.6 +/- 0.5 mM, 100-fold higher than that for Cel 7A acting on low-molecular-weight model substrates. The hydrolysis of [(3)H]-amorphous cellulose by endoglucanases was even less affected by cellobiose inhibition with apparent K(i) values of 11 +/- 3 mM and 34 +/- 6 mM for Cel 7B and Cel 5A, respectively. Contrary to the case for the other enzymes studied, the release of Radioactive Label by Cel 12A was stimulated by cellobiose, possibly due to a more pronounced transglycosylating activity. Theoretical analysis of the inhibition of Cel 7A by cellobiose predicted an inhibition analogous to that of mixed type with two limiting cases, competitive inhibition if the prevalent enzyme-substrate complex without inhibitor is productive and conventional mixed type when the prevalent enzyme-substrate complex is nonproductive.

  • inhibition of the trichoderma reesei cellulases by cellobiose is strongly dependent on the nature of the substrate
    Biotechnology and Bioengineering, 2004
    Co-Authors: Marju Gruno, Priit Valjamae, Goran Pettersson, Gunnar Johansson
    Abstract:

    The inhibition effect of cellobiose on the initial stage of hydrolysis when cellobiohydrolase Cel 7A and endoglucanases Cel 7B, Cel 5A, and Cel 12A from Trichoderma reesei were acting on bacterial cellulose and amorphous cellulose that were [3H]- Labeled at the reducing end was quantified. The apparent competitive inhibition constant (Ki) for Cel 7A on [3H]-bacterial cellulose was found to be 1.6 ± 0.5 mM, 100-fold higher than that for Cel 7A acting on low-molecular-weight model substrates. The hydrolysis of [3H]-amorphous cellulose by endoglucanases was even less affected by cellobiose inhibition with apparent Ki values of 11 ± 3 mM and 34 ± 6 mM for Cel 7B and Cel 5A, respectively. Contrary to the case for the other enzymes studied, the release of Radioactive Label by Cel 12A was stimulated by cellobiose, possibly due to a more pronounced transglycosylating activity. Theoretical analysis of the inhibition of Cel 7A by cellobiose predicted an inhibition analogous to that of mixed type with two limiting cases, competitive inhibition if the prevalent enzyme-substrate complex without inhibitor is productive and conventional mixed type when the prevalent enzyme-substrate complex is nonproductive. © 2004 Wiley Periodicals, Inc.

Bryce P Nelson - One of the best experts on this subject based on the ideXlab platform.

  • Label free detection of 16s ribosomal rna hybridization on reusable dna arrays using surface plasmon resonance imaging
    Environmental Microbiology, 2002
    Co-Authors: Bryce P Nelson, Mark R Liles, Kendra B Frederick, Robert M Corn, Robert M Goodman
    Abstract:

    Summary In this paper, we describe the detection of bacterial cell-extracted 16S ribosomal RNA (rRNA) using an emerging technology, surface plasmon resonance (SPR) imaging of DNA arrays. Surface plasmon reso- nance enables detection of molecular interactions on surfaces in response to changes in the index of refraction, therefore eliminating the need for a fluo- rescent or Radioactive Label. A variation of the more common SPR techniques, SPR imaging enables detection from multiple probes in a reusable array format. The arrays developed here contain DNA probes (15-21 bases) designed to be complementary to 16S rRNA gene sequences of Escherichia coli and Bacillus subtilis as well as to a highly conserved sequence found in rRNAs from most members of the domain Bacteria. We report species-specific hybrid- ization of cell-extracted total RNA and in vitro tran- scribed 16S rRNA to oligonucleotide probes on SPR arrays. We tested multiple probe sequences for each species, and found that success or failure of hybrid- ization was dependent upon probe position in the 16S rRNA molecule. It was also determined that one of the probes intended to bind 16S rRNA also bound an unknown protein. The amount of binding to these probes was quantified with SPR imaging. A detection limit of 2 m g ml - - - 1 was determined for fragmented E.

  • Label free detection of 16s ribosomal rna hybridization on reusable dna arrays using surface plasmon resonance imaging
    Environmental Microbiology, 2002
    Co-Authors: Bryce P Nelson, Mark R Liles, Kendra B Frederick, Robert M Corn, Robert M Goodman
    Abstract:

    In this paper, we describe the detection of bacterial cell-extracted 16S ribosomal RNA (rRNA) using an emerging technology, surface plasmon resonance (SPR) imaging of DNA arrays. Surface plasmon resonance enables detection of molecular interactions on surfaces in response to changes in the index of refraction, therefore eliminating the need for a fluorescent or Radioactive Label. A variation of the more common SPR techniques, SPR imaging enables detection from multiple probes in a reusable array format. The arrays developed here contain DNA probes (15-21 bases) designed to be complementary to 16S rRNA gene sequences of Escherichia coli and Bacillus subtilis as well as to a highly conserved sequence found in rRNAs from most members of the domain Bacteria. We report species-specific hybridization of cell-extracted total RNA and in vitro transcribed 16S rRNA to oligonucleotide probes on SPR arrays. We tested multiple probe sequences for each species, and found that success or failure of hybridization was dependent upon probe position in the 16S rRNA molecule. It was also determined that one of the probes intended to bind 16S rRNA also bound an unknown protein. The amount of binding to these probes was quantified with SPR imaging. A detection limit of 2 micro g ml-1 was determined for fragmented E. coli total cellular RNA under the experimental conditions used. These results indicate the feasibility of using SPR imaging for 16S rRNA identification and encourage further development of this method for direct detection of other RNA molecules.

Priit Valjamae - One of the best experts on this subject based on the ideXlab platform.

  • inhibition of the trichoderma reesei cellulases by cellobiose is strongly dependent on the nature of the substrate
    Biotechnology and Bioengineering, 2004
    Co-Authors: Marju Gruno, Priit Valjamae, Goran Pettersson, Gunnar Johansson
    Abstract:

    The inhibition effect of cellobiose on the initial stage of hydrolysis when cellobiohydrolase Cel 7A and endoglucanases Cel 7B, Cel 5A, and Cel 12A from Trichoderma reesei were acting on bacterial cellulose and amorphous cellulose that were [(3)H]- Labeled at the reducing end was quantified. The apparent competitive inhibition constant (K(i)) for Cel 7A on [(3)H]-bacterial cellulose was found to be 1.6 +/- 0.5 mM, 100-fold higher than that for Cel 7A acting on low-molecular-weight model substrates. The hydrolysis of [(3)H]-amorphous cellulose by endoglucanases was even less affected by cellobiose inhibition with apparent K(i) values of 11 +/- 3 mM and 34 +/- 6 mM for Cel 7B and Cel 5A, respectively. Contrary to the case for the other enzymes studied, the release of Radioactive Label by Cel 12A was stimulated by cellobiose, possibly due to a more pronounced transglycosylating activity. Theoretical analysis of the inhibition of Cel 7A by cellobiose predicted an inhibition analogous to that of mixed type with two limiting cases, competitive inhibition if the prevalent enzyme-substrate complex without inhibitor is productive and conventional mixed type when the prevalent enzyme-substrate complex is nonproductive.

  • inhibition of the trichoderma reesei cellulases by cellobiose is strongly dependent on the nature of the substrate
    Biotechnology and Bioengineering, 2004
    Co-Authors: Marju Gruno, Priit Valjamae, Goran Pettersson, Gunnar Johansson
    Abstract:

    The inhibition effect of cellobiose on the initial stage of hydrolysis when cellobiohydrolase Cel 7A and endoglucanases Cel 7B, Cel 5A, and Cel 12A from Trichoderma reesei were acting on bacterial cellulose and amorphous cellulose that were [3H]- Labeled at the reducing end was quantified. The apparent competitive inhibition constant (Ki) for Cel 7A on [3H]-bacterial cellulose was found to be 1.6 ± 0.5 mM, 100-fold higher than that for Cel 7A acting on low-molecular-weight model substrates. The hydrolysis of [3H]-amorphous cellulose by endoglucanases was even less affected by cellobiose inhibition with apparent Ki values of 11 ± 3 mM and 34 ± 6 mM for Cel 7B and Cel 5A, respectively. Contrary to the case for the other enzymes studied, the release of Radioactive Label by Cel 12A was stimulated by cellobiose, possibly due to a more pronounced transglycosylating activity. Theoretical analysis of the inhibition of Cel 7A by cellobiose predicted an inhibition analogous to that of mixed type with two limiting cases, competitive inhibition if the prevalent enzyme-substrate complex without inhibitor is productive and conventional mixed type when the prevalent enzyme-substrate complex is nonproductive. © 2004 Wiley Periodicals, Inc.

Mark R Liles - One of the best experts on this subject based on the ideXlab platform.

  • Label free detection of 16s ribosomal rna hybridization on reusable dna arrays using surface plasmon resonance imaging
    Environmental Microbiology, 2002
    Co-Authors: Bryce P Nelson, Mark R Liles, Kendra B Frederick, Robert M Corn, Robert M Goodman
    Abstract:

    Summary In this paper, we describe the detection of bacterial cell-extracted 16S ribosomal RNA (rRNA) using an emerging technology, surface plasmon resonance (SPR) imaging of DNA arrays. Surface plasmon reso- nance enables detection of molecular interactions on surfaces in response to changes in the index of refraction, therefore eliminating the need for a fluo- rescent or Radioactive Label. A variation of the more common SPR techniques, SPR imaging enables detection from multiple probes in a reusable array format. The arrays developed here contain DNA probes (15-21 bases) designed to be complementary to 16S rRNA gene sequences of Escherichia coli and Bacillus subtilis as well as to a highly conserved sequence found in rRNAs from most members of the domain Bacteria. We report species-specific hybrid- ization of cell-extracted total RNA and in vitro tran- scribed 16S rRNA to oligonucleotide probes on SPR arrays. We tested multiple probe sequences for each species, and found that success or failure of hybrid- ization was dependent upon probe position in the 16S rRNA molecule. It was also determined that one of the probes intended to bind 16S rRNA also bound an unknown protein. The amount of binding to these probes was quantified with SPR imaging. A detection limit of 2 m g ml - - - 1 was determined for fragmented E.

  • Label free detection of 16s ribosomal rna hybridization on reusable dna arrays using surface plasmon resonance imaging
    Environmental Microbiology, 2002
    Co-Authors: Bryce P Nelson, Mark R Liles, Kendra B Frederick, Robert M Corn, Robert M Goodman
    Abstract:

    In this paper, we describe the detection of bacterial cell-extracted 16S ribosomal RNA (rRNA) using an emerging technology, surface plasmon resonance (SPR) imaging of DNA arrays. Surface plasmon resonance enables detection of molecular interactions on surfaces in response to changes in the index of refraction, therefore eliminating the need for a fluorescent or Radioactive Label. A variation of the more common SPR techniques, SPR imaging enables detection from multiple probes in a reusable array format. The arrays developed here contain DNA probes (15-21 bases) designed to be complementary to 16S rRNA gene sequences of Escherichia coli and Bacillus subtilis as well as to a highly conserved sequence found in rRNAs from most members of the domain Bacteria. We report species-specific hybridization of cell-extracted total RNA and in vitro transcribed 16S rRNA to oligonucleotide probes on SPR arrays. We tested multiple probe sequences for each species, and found that success or failure of hybridization was dependent upon probe position in the 16S rRNA molecule. It was also determined that one of the probes intended to bind 16S rRNA also bound an unknown protein. The amount of binding to these probes was quantified with SPR imaging. A detection limit of 2 micro g ml-1 was determined for fragmented E. coli total cellular RNA under the experimental conditions used. These results indicate the feasibility of using SPR imaging for 16S rRNA identification and encourage further development of this method for direct detection of other RNA molecules.