title: Chloroplast-Based Expression of Recombinant Proteins by Gateway® Cloning Technology.
abstract: Plastid transformation for the expression of recombinant proteins and entire enzymatic pathways has become a promising tool for plant biotechnology in the past decade. Several improvements of the technology have turned plant plastids into robust and dependable expression platforms for multiple high value compounds. In this chapter, we describe our current methodology based on Gateway(®) recombinant cloning, which we have adapted for plastid transformation. We describe the steps required for cloning, biolistic transformation, identification, and regeneration of transplastomic plant lines and Western blot analysis.
authors.affiliation: Department of Crop Sciences, University of Natural Resources and Life Sciences, Konrad Lorenz Str. 24, A - 3430, Tulln a. d. Donau, Austria.
authors.forename: Johanna
authors.initials: J
authors.lastname: Gottschamel
authors.affiliation: Department of Crop Sciences, University of Natural Resources and Life Sciences, Konrad Lorenz Str. 24, A - 3430, Tulln a. d. Donau, Austria. andreas.loessl@boku.ac.at.
authors.forename: Andreas
authors.initials: A
authors.lastname: Lössl
mesh_terms: D019470:Biolistics; D015153:Blotting, Western; D002736:Chloroplasts / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D030821:Plants, Genetically Modified; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic
chemical_list: D011994:Recombinant Proteins
keywords: Biolistic transformation; Gateway® recombinant cloning technology; Plastid transformation; Southern blot analysis; Western blot analysis