The curves showing expression profiles of all other genes of the ATP synthase operon are in gray. Microarray values were background-corrected, normalized against the median of the ratio of each sample against the reference, and log-transformed. The plotted data include microarray replicates of 38 biological experiments. b The arrangement of genes of the ATP synthase operon. The genes are depicted as arrows, with the orientation indicated by the direction of the arrow. The location of the genes on the chromosome relative to the origin is indicated. This information
was obtained from CyanoBase (http://genome.kazusa.or.jp/cyanobase/) (Nakao et al. 2010). The genes of the operon are atp1 (sll1321), atpI (sll1322), atpH (ssl2615), atpG (sll1323), atpF (sll1324), atpD (sll1325), atpA (sll1326), and atpC (sll1327). slr1413 Mizoribine is upstream, and slr1411 and sll0216 are downstream of the ATP synthase operon, respectively, and neither is co-expressed with atp1. All of the genes of the ATP synthase operon are depicted as light gray-filled arrows, except for atp1; this arrow is red-filled. NVP-BEZ235 supplier Arrows representing genes outside the operon, slr1411, slr1413, and sll0216, are unfilled and dark gray-filled Phenotypic analysis of GreenCut mutants Identification of numerous proteins potentially involved in photosynthetic function
allows for the exploitation of reverse genetic approaches to generate specific strains Bay 11-7085 that are null or suppressed for a specific targeted gene. Strategies that have been successfully used to generate such strains include RNAi (Rohr et al. 2004; Im et al. 2006) and amiRNA approaches (Molnar et al. 2009; Zhao et al. 2009), as well as PCR identification of strains harboring specific mutations (Pootakham et al. 2010). Thus far, approximately 30 strains of Chlamydomonas and well over 100 strains of Arabidopsis have been identified with insertions in genes encoding GreenCut proteins of unknown function. Both sets of mutants are
being analyzed using a specific set of assays that are relatively rapid. An example of a specific Chlamydomonas mutant strain that has gone through the primary assays of the characterization platform potentially harbors a lesion in the gene encoding CGL28, which has a motif that may allow it to bind RNA. Initially, the cells are grown on both minimal medium (no fixed carbon source) supplemented with bicarbonate and medium containing acetate. As shown in Fig. 3, a Chlamydomonas strain with a lesion in CGL28 (colony within red box, step 1) appears to be unable to grow on minimal medium, although it can grow on medium supplemented with acetate. The colonies that grew on acetate-containing medium were examined for fluorescence to determine the quantum yield of PSII. The fluorescence image shown in Fig.