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  • Invasion assay  (1)
  • Mutagenesis  (1)
  • Springer  (2)
  • 1
    Electronic Resource
    Electronic Resource
    Site-specific homologous recombination mutagenesis in group B streptococci (1998)
    Springer
    Methods in cell science 20 (1998), S. 13-20 
    Details
    ISSN: 1573-0603
    Keywords: Capsule synthesis ; Mutagenesis ; Recombination ; Streptococci ; Suicide vector
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We describe the use of suicide vectors to generate site-specific mutations in group B streptococcus. This is accomplished by cloning gene-specific sequences into a temperature sensitive plasmid and selecting for clones which have undergone homologous recombination. The recombinan clones can be easily isolated by selecting for clones which have retained the antibiotic resistance markers that are present on the vector or cloned into the gene-specific sequences. To confirm the fidelity of the recombination events, PCR analysis is performed on chromosomal DNA isolated from the recombinant clones. Using this strategy, we have generated site- specific insertions in several capsule genes and have found that these insertions occurred as expected and that the mutations result in loss of capsule expression. In this report, we specifically detail the construction of cpsB mutants by single and double cross-over recombination and demonstrate that the resulting strains are acapsular.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009810002276
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  • 2
    Electronic Resource
    Electronic Resource
    A simple microtiter plate screening assay for bacterial invasion or adherence (1998)
    Springer
    Methods in cell science 20 (1998), S. 107-111 
    Details
    ISSN: 1573-0603
    Keywords: Bacterial adherence ; Bacterial invasion ; Bacteriological techniques ; Invasion assay ; Microbial colony count ; Soft agar
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Many bacteriologic studies, including cellular invasion and adherence assays, require enumeration of viable organisms or colony forming units. When attempting to screen large numbers of clinical or environmental isolates or laboratory-derived mutants for differences in invasion or adherence phenotype, standard plating methods can be cumbersome and severely limit the number of organisms or conditions which can be tested. As a potential alternative, we describe a simple, rapid and inexpensive soft agar-based technique for semi-quantitative determination of bacterial colony counts directly within the wells of a 96-well microtiter plate.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009830606819
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