Abstract
Microinjection of mammalian cells provides a powerful method for analyzing in vivo functions of viral genes and viral gene products. By microinjection, a controlled amount (ranging from several to many thousands of copies) of a viral or cellular gene, a protein product of a gene, a polypeptide fragment encoding a specific protein domain, or an RNA molecule can be delivered into a target cell and the functional consequences analyzed. Microinjection can be used to deliver antibody targeted to a specific protein domain in order to analyze the requirement of the protein for specific cell functions such as cell cycle progression, transcription of specific genes, or intracellular transport.
This chapter describes examples of the successful use of microinjection to probe adenovirus E1A regulatory mechanisms. Detailed methods are provided for manual and semiautomatic microinjection of mammalian cells as well as bioassay protocols for microinjected cells including immunofluorescence, colorimetic, in situ hybridization, and autoradiography.
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© 2007 Humana Press Inc, Totowa, NJ
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Green, M., Thorburn, A., Kern, R., Loewenstein, P.M. (2007). The Use of Cell Microinjection for the In Vivo Analysis of Viral Transcriptional Regulatory Protein Domains. In: Wold, W.S.M., Tollefson, A.E. (eds) Adenovirus Methods and Protocols. Methods in Molecular Medicine™, vol 131. Humana Press. https://doi.org/10.1007/978-1-59745-277-9_12
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DOI: https://doi.org/10.1007/978-1-59745-277-9_12
Publisher Name: Humana Press
Print ISBN: 978-1-58829-901-7
Online ISBN: 978-1-59745-277-9
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