Berliner Und Munchener Tierarztliche Wochenschrift, 2003
Microarrays (DNA-Chips) are miniaturized carriers on which many nucleic acid molecule probes such... more Microarrays (DNA-Chips) are miniaturized carriers on which many nucleic acid molecule probes such as oligonucleotides or PCR products are immobilized in a high density, and compactness. Homologue DNA hybridises with the immobilized complementary nucleic acid probes. This study gives after a short general introduction in the principle of DNA-microarrays an overview about published data on the field of typing of Salmonella by microarrays. An onset of a DNA-microarray developed by the National Reference Laboratory for Salmonella (NRL-Salm) will be introduced. By this new technique, it is possible to answer epidemiological questions as well as to find genes involved in certain biochemical processes, such as pathogenicity or resistance of salmonellae.
Molekularbiologische Methoden in der Lebensmittelanalytik, 2010
Mit der Verbreitung des Begriffes „Biochip“ in den biotechnologischen Medien wurde Ende der 1990e... more Mit der Verbreitung des Begriffes „Biochip“ in den biotechnologischen Medien wurde Ende der 1990er-Jahre zunächst der Eindruck erweckt, dass die Computerelektronik in die molekularbiologischen Anwendungen eingestiegen ist [18]. In nur wenigen Jahren hat sich die Biochiptechnologie zu einem Verfahren entwickelt, das aus der molekularbiologischen Grundlagenforschung nicht mehr wegzudenken ist und über eine Vielzahl von Einsatzbereichen verfügt. Die Biochiptechnologie ermöglicht die
Berliner und Münchener tierärztliche Wochenschrift
Microarrays (DNA-Chips) are miniaturized carriers on which many nucleic acid molecule probes such... more Microarrays (DNA-Chips) are miniaturized carriers on which many nucleic acid molecule probes such as oligonucleotides or PCR products are immobilized in a high density, and compactness. Homologue DNA hybridises with the immobilized complementary nucleic acid probes. This study gives after a short general introduction in the principle of DNA-microarrays an overview about published data on the field of typing of Salmonella by microarrays. An onset of a DNA-microarray developed by the National Reference Laboratory for Salmonella (NRL-Salm) will be introduced. By this new technique, it is possible to answer epidemiological questions as well as to find genes involved in certain biochemical processes, such as pathogenicity or resistance of salmonellae.
MxA is a key component in the interferon-induced antiviral defense in humans. After viral infecti... more MxA is a key component in the interferon-induced antiviral defense in humans. After viral infections, MxA is rapidly induced and accumulates in the cytoplasm. The multiplication of many RNA viruses, including all bunyaviruses tested so far, is inhibited by MxA. These findings prompted us to express MxA in plants in an attempt to create resistance to tospoviruses. Here, we report the generation of transgenic tobacco plants that constitutively express MxA under the control of the 35 S cauliflower mosaic virus promotor. Northern and western blot analysis confirmed the expression of MxA in several transgenic plant lines. MxA expression had no obvious detrimental effects on plant growth and fertility. However, challenge experiments with tomato spotted wilt virus, tomato chlorotic spot virus, and groundnut ringspot virus revealed no increased resistance of MxA-transgenic tobacco plants to tospovirus infections. Neither was the multiplication of tobacco mosaic virus, cucumber mosaic virus ...
Sequence analysis of amplified cDNAs derived from the maize chloroplast rpoB transcript which enc... more Sequence analysis of amplified cDNAs derived from the maize chloroplast rpoB transcript which encodes the beta subunit of a chloroplast specific, DNA dependent RNA polymerase reveals four C-to-U editing sites clustered within 150 nucleotides of the 5' terminal region of the rpoB message. These newly identified editing sites confirm the bias of chloroplast editing for certain codon transitions and for second codon positions which both appear suggestive for an involvement of the translational apparatus in the editing process. This supposition prompted us to investigate editing of the rpoB transcript from ribosome deficient, and hence protein synthesis deficient, plastids of the barley mutant albostrians. In this mutant editing is, however, not impaired at any of the editing sites functional in the barley wild type rpoB transcript. This demonstrates that chloroplast editing is neither linked to nor dependent on the chloroplast translational apparatus. As a further consequence any p...
The NADH dehydrogenase subunit A (ndhA) gene from maize chloroplasts encodes a highly conserved p... more The NADH dehydrogenase subunit A (ndhA) gene from maize chloroplasts encodes a highly conserved peptide, which at several positions could be restored to consensus sequences by potential C-to4 editing of the codons involved. This gene was, therefore, chosen for analysis of its mRNA sequence in the form of amplified cDNA. A comparison of this cDNA sequence with the plastome-encoded ndhA
Plastid RNA editing of a number of transcripts at specific sites changes genomically encoded cyti... more Plastid RNA editing of a number of transcripts at specific sites changes genomically encoded cytidines to nucleosides, which act like uridines in RT-PCR analyses. To study plastid-editing directly at the RNA level, we established a single-strand conformational polymorphism assay for the discrimination of small RNA molecules. The electrophoretic mobility of a oligoribonucleotide resulting from a RNase T1-digested and edited plastid mRNA was shown to be identical with a control RNA molecule containing a uridine at the editing site, whereas the unedited RNA behaved like a RNA molecule containing a cytidine at the respective position.
Berliner Und Munchener Tierarztliche Wochenschrift, 2003
Microarrays (DNA-Chips) are miniaturized carriers on which many nucleic acid molecule probes such... more Microarrays (DNA-Chips) are miniaturized carriers on which many nucleic acid molecule probes such as oligonucleotides or PCR products are immobilized in a high density, and compactness. Homologue DNA hybridises with the immobilized complementary nucleic acid probes. This study gives after a short general introduction in the principle of DNA-microarrays an overview about published data on the field of typing of Salmonella by microarrays. An onset of a DNA-microarray developed by the National Reference Laboratory for Salmonella (NRL-Salm) will be introduced. By this new technique, it is possible to answer epidemiological questions as well as to find genes involved in certain biochemical processes, such as pathogenicity or resistance of salmonellae.
Molekularbiologische Methoden in der Lebensmittelanalytik, 2010
Mit der Verbreitung des Begriffes „Biochip“ in den biotechnologischen Medien wurde Ende der 1990e... more Mit der Verbreitung des Begriffes „Biochip“ in den biotechnologischen Medien wurde Ende der 1990er-Jahre zunächst der Eindruck erweckt, dass die Computerelektronik in die molekularbiologischen Anwendungen eingestiegen ist [18]. In nur wenigen Jahren hat sich die Biochiptechnologie zu einem Verfahren entwickelt, das aus der molekularbiologischen Grundlagenforschung nicht mehr wegzudenken ist und über eine Vielzahl von Einsatzbereichen verfügt. Die Biochiptechnologie ermöglicht die
Berliner und Münchener tierärztliche Wochenschrift
Microarrays (DNA-Chips) are miniaturized carriers on which many nucleic acid molecule probes such... more Microarrays (DNA-Chips) are miniaturized carriers on which many nucleic acid molecule probes such as oligonucleotides or PCR products are immobilized in a high density, and compactness. Homologue DNA hybridises with the immobilized complementary nucleic acid probes. This study gives after a short general introduction in the principle of DNA-microarrays an overview about published data on the field of typing of Salmonella by microarrays. An onset of a DNA-microarray developed by the National Reference Laboratory for Salmonella (NRL-Salm) will be introduced. By this new technique, it is possible to answer epidemiological questions as well as to find genes involved in certain biochemical processes, such as pathogenicity or resistance of salmonellae.
MxA is a key component in the interferon-induced antiviral defense in humans. After viral infecti... more MxA is a key component in the interferon-induced antiviral defense in humans. After viral infections, MxA is rapidly induced and accumulates in the cytoplasm. The multiplication of many RNA viruses, including all bunyaviruses tested so far, is inhibited by MxA. These findings prompted us to express MxA in plants in an attempt to create resistance to tospoviruses. Here, we report the generation of transgenic tobacco plants that constitutively express MxA under the control of the 35 S cauliflower mosaic virus promotor. Northern and western blot analysis confirmed the expression of MxA in several transgenic plant lines. MxA expression had no obvious detrimental effects on plant growth and fertility. However, challenge experiments with tomato spotted wilt virus, tomato chlorotic spot virus, and groundnut ringspot virus revealed no increased resistance of MxA-transgenic tobacco plants to tospovirus infections. Neither was the multiplication of tobacco mosaic virus, cucumber mosaic virus ...
Sequence analysis of amplified cDNAs derived from the maize chloroplast rpoB transcript which enc... more Sequence analysis of amplified cDNAs derived from the maize chloroplast rpoB transcript which encodes the beta subunit of a chloroplast specific, DNA dependent RNA polymerase reveals four C-to-U editing sites clustered within 150 nucleotides of the 5' terminal region of the rpoB message. These newly identified editing sites confirm the bias of chloroplast editing for certain codon transitions and for second codon positions which both appear suggestive for an involvement of the translational apparatus in the editing process. This supposition prompted us to investigate editing of the rpoB transcript from ribosome deficient, and hence protein synthesis deficient, plastids of the barley mutant albostrians. In this mutant editing is, however, not impaired at any of the editing sites functional in the barley wild type rpoB transcript. This demonstrates that chloroplast editing is neither linked to nor dependent on the chloroplast translational apparatus. As a further consequence any p...
The NADH dehydrogenase subunit A (ndhA) gene from maize chloroplasts encodes a highly conserved p... more The NADH dehydrogenase subunit A (ndhA) gene from maize chloroplasts encodes a highly conserved peptide, which at several positions could be restored to consensus sequences by potential C-to4 editing of the codons involved. This gene was, therefore, chosen for analysis of its mRNA sequence in the form of amplified cDNA. A comparison of this cDNA sequence with the plastome-encoded ndhA
Plastid RNA editing of a number of transcripts at specific sites changes genomically encoded cyti... more Plastid RNA editing of a number of transcripts at specific sites changes genomically encoded cytidines to nucleosides, which act like uridines in RT-PCR analyses. To study plastid-editing directly at the RNA level, we established a single-strand conformational polymorphism assay for the discrimination of small RNA molecules. The electrophoretic mobility of a oligoribonucleotide resulting from a RNase T1-digested and edited plastid mRNA was shown to be identical with a control RNA molecule containing a uridine at the editing site, whereas the unedited RNA behaved like a RNA molecule containing a cytidine at the respective position.
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