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An Improved Transient Expression System Using Arabidopsis Protoplasts

Tipo de material: TextoTextoSeries ; Current Protocols in Plant Biology, 1, p.285-291, 2016Trabajos contenidos:
  • Cao, Yangrong
  • Li, Hong
  • Pham, An Q
Tema(s): Recursos en línea: Resumen: Transient gene expression in protoplasts provides a powerful tool to study protein expression, protein localization, protein-protein association, and gene expression regulation, etc. There are several methods including electroporation, which have been reported to introduce DNA into protoplasts. However, one of the best methods used is polyethylene glycol (PEG)-mediated transfection. Here, we describe an improved PEG-mediated transformation method including preparation of protoplasts, PEG-mediated transformation, and, by way of example, expression of the AtLYK5 gene (AT2G33580)in protoplasts. The protoplast transient expression system provides unique capabilities to support cell-based experiments involved in plant biochemistry and physiology.
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Transient gene expression in protoplasts provides a powerful tool to study protein expression, protein localization, protein-protein association, and gene expression regulation, etc. There are several methods including electroporation, which have been reported to introduce DNA into protoplasts. However, one of the best methods used is polyethylene glycol (PEG)-mediated transfection. Here, we describe an improved PEG-mediated transformation method including preparation of protoplasts, PEG-mediated transformation, and, by way of example, expression of the AtLYK5 gene (AT2G33580)in protoplasts. The protoplast transient expression system provides unique capabilities to support cell-based experiments involved in plant biochemistry and physiology.

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