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  • 11
    Electronic Resource
    Electronic Resource
    Improved detection of prunus necrotic ringspot virus by the polymerase chain reaction (1996)
    Springer
    European journal of plant pathology 102 (1996), S. 681-685 
    Details
    ISSN: 1573-8469
    Keywords: Ilarvirus ; PCR ; peach ; PNRSV ; virus detection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The reverse transcription — polymerase chain reaction (RT-PCR) technique was used for detection of prunus necrotic ringspot virus in dormant peach trees which tested negative by ELISA. Total RNA extracted from bark tissue, using a lithium chloride based method, were used for reverse transcription and subsequent amplification of viral sequences. The PCR product, about 300 base pairs in size, was analyzed by gel electrophoresis and visualized by ethidium bromide staining. In some cases, PCR products were not clearly visible in the stained gel and became distinct only following hybridisation with a32P-labelled virus specific probe. The RT-PCR assay described in this paper is sensitive enough for detection of PNRSV in dormant woody bark tissue and could be incorporated into testing protocols during post-entry quarantines for rapid initial screening of imported budwood and in virus elimination programs.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01877249
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    Paper (German National Licenses)
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  • 12
    Electronic Resource
    Electronic Resource
    Heterogeneity of the genes coding for 5 S RNA in three related strains of the genus Bacillus (1977)
    Springer
    Molecular genetics and genomics 156 (1977), S. 185-193 
    Details
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Three related strains of the genus Bacillus, viz. B. licheniformis, B. subtilis and Bacillus Q were all found to contain a minor species of 5 S RNA in an amount indicating that it is transcribed from only one of the multiple 5 S RNA cistrons known to be present in these strains. The major and minor types of 5 S RNA from each of the three strains could be separated from each other by polyacrylamide gel electrophoresis in the presence of urea. The complete nucleotide sequences of the minor B. subtilis and Bacillus Q 5 S RNAs have been determined. Comparison of these sequences to the previously determined sequence of the minor 5 S RNA from B. licheniformis (Raué et al., 1976) showed the three minor types of 5 S RNA to be identical except for the residues at positions 79, 85 and 95. Moreover, seven out of the eight sequence differences between the major and the minor 5 S RNA are the same in all three strains. Thus, the single cistron coding for minor 5 S RNA has been strongly conserved in all three strains, which may indicate a biological significance for the minor 5 S RNA species.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00283491
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    Paper (German National Licenses)
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  • 13
    Electronic Resource
    Electronic Resource
    Use of the polymerase chain reaction to clone the potato leafroll virus coat protein gene directly from the total RNA of infected plants (1995)
    Springer
    Potato research 38 (1995), S. 211-218 
    Details
    ISSN: 1871-4528
    Keywords: reverse transcription ; PCR. DNA sequencing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The potato leafroll virus (PLRV) coat protein (CP) gene was directly cloned from the total RNA extracted from virus-infected plants. First strand cDNA synthesis was not necessarily specific; it was equally efficient using either random or CP-specific primers. The viral sequence encoding the coat protein was specifically amplified from the total population of cDNA molecules by polymerase chain reaction (PCR), using specific primers bordering the CP gene. The unique amplified product thus obtained was cloned blunt-end into the pT7T318U plasmid vector, and the authenticity of the cloned gene verified by sequence analysis. This cloning strategy obviates the need for virus purification. Sequence comparison of the CP gene of the Italian isolate and those of five other PLRV isolates revealed a close similarity to the three European and the Canadian isolates, and a more distant relationship with the Australian one.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02357934
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    Paper (German National Licenses)
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