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Reverse Transfected Cell Microarrays in Infectious Disease Research

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Cell-Based Microarrays

Part of the book series: Methods in Molecular Biology ((MIMB,volume 706))

Abstract

Several human pathogenic viruses encode large genomes with often more than 100 genes. Viral pathogenicity is determined by carefully orchestrated co-operative activities of several different viral genes which trigger the phenotypic functions of the infected cells. Systematic analyses of these complex interactions require high-throughput transfection technology. Here we have provided a laboratory manual for the reverse transfected cell microarray (RTCM; alternative name: cell chip) as a high-throughput transfection procedure, which has been successfully applied for the systematic analyses of single and combination effects of genes encoded by the human herpesvirus-8 on the NF-kappaB signal transduction pathway. In order to quantitatively determine the effects of viral genes in transfected cells, protocols for the use of GFP as an indicator gene and for indirect immunofluorescence staining of cellular target proteins have been included. RTCM provides a useful methodological approach to investigate systematically combination effects of viral genes on cellular functions.

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Acknowledgments

This work was supported by grants of the Deutsche Forschungsgemeinschaft (DFG-SPP 1130, DFG-GK 1071, and DFG 317/2-1) and of the Interdisciplinary Center for Clinical Research (IZKF) of the University Medical Center Erlangen to MS and by a habilitation grant (Habilitationsstipendium, ELAN-Förderprogramm) of the University Medical Center Erlangen to RJ.

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Authors and Affiliations

  1. Division of Molecular and Experimental Surgery, Department of Surgery, University Medical Center Erlangen, Erlangen, Germany

    Andreas Konrad, Ramona Jochmann, Elisabeth Kuhn, Elisabeth Naschberger, Priya Chudasama & Michael Stürzl

Authors
  1. Andreas Konrad
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  2. Ramona Jochmann
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  3. Elisabeth Kuhn
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  4. Elisabeth Naschberger
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  5. Priya Chudasama
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  6. Michael Stürzl
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Corresponding author

Correspondence to Andreas Konrad .

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Editors and Affiliations

  1. MRC Clinical Sciences Centre, Faculty of Medicine, Imperial College London, Du Cane Road, London, W12 0NN, United Kingdom

    Ella Palmer

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Konrad, A., Jochmann, R., Kuhn, E., Naschberger, E., Chudasama, P., Stürzl, M. (2011). Reverse Transfected Cell Microarrays in Infectious Disease Research. In: Palmer, E. (eds) Cell-Based Microarrays. Methods in Molecular Biology, vol 706. Humana Press. https://doi.org/10.1007/978-1-61737-970-3_9

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  • DOI: https://doi.org/10.1007/978-1-61737-970-3_9

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61737-969-7

  • Online ISBN: 978-1-61737-970-3

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