title: Rapid screening for the robust expression of recombinant proteins in algal plastids.
abstract: Chlamydomonas reinhardtii has many advantages as a photosynthetic model organism. One of these is facile, targeted chloroplast transformation by particle bombardment. Functional recombinant proteins can be expressed to significant levels in this system, potentially outperforming higher plants in speed of scaling, cost, and space requirements. Several strategies and regulatory regions can be used for achieving transgene expression. Here we present two of those strategies: one makes use of the psbD promoter for expressing moderate levels of the recombinant protein in a photosynthetic background. The other strategy is based on the strong psbA promoter for obtaining high yields of the recombinant product in a non-photosynthetic strain. We herein describe the vectors, transformation procedures, and screening methods associated with these two strategies.
authors.affiliation: California Center for Algae Biotechnology, La Jolla, CA, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Barrera
authors.forename: Javier
authors.initials: J
authors.lastname: Gimpel
authors.forename: Stephen
authors.initials: S
authors.lastname: Mayfield
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D019470:Biolistics / Q000379:methods; D016825:Chlamydomonas reinhardtii / Q000235:genetics*; D002736:Chloroplasts / Q000235:genetics* / Q000378:metabolism; D004351:Drug Resistance / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D007612:Kanamycin / Q000494:pharmacology; D010788:Photosynthesis / Q000235:genetics*; D045332:Photosystem II Protein Complex / Q000235:genetics*; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D021122:Protein Subunits / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics*; D019076:Transgenes / Q000235:genetics
chemical_list: D000900:Anti-Bacterial Agents; D045332:Photosystem II Protein Complex; D021122:Protein Subunits; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D007612:Kanamycin