Poster: Photosynthesis

Abs # 346: Chloroplast transformation involving mutations in the atpE gene in the green alga Chlamydomonas reinhardtii

Presenter:	Johnson, Eric , ejohn@nature.berkeley.edu
Authors	Johnson, Eric  (A)   Melils, Anastasios  (A)
Affiliations:	(A): University of California

The atpE gene is localized on the chloroplast genome in Chlamydomonas reinhardtii and codes for the epsilon subunit of the chloroplast ATP synthase. A strain of Chlamydomonas reinhardtii carrying a defective version of the atpE gene (FUD 17) has been used in biolistic transformations, receiving a plasmid, pUCatpE, which contained the wild type atpE gene. This transformed strain regains the ability for photo-autotrophic growth. A mutation was introduced into pUCatpE, using PCR based mutagenesis, which alters the atpE gene incorporating a stop codon six amino acids before the wild type stop codon. This new plasmid was used in biolistic transformations of the FUD17. A new strain of C. reinhardtii was generated, which also regained photosynthetic growth and expressed an ATP synthase with a truncated epsilon subunit. This biolistic transformation technique will be used to generate mutations of the epsilon subunit and to study the function and assembly of chloroplast ATP synthase in vivo.