Functional Analysis of a Rice Oxidosqualene Cyclase through Total Gene Synthesis  Juncong Sun, Xia Xu, Zheyong Xue, John Hugh Snyder, Xiaoquan Qi  Molecular Plant  Volume 6, Issue 5, Pages 1726-1729 (September 2013) DOI: 10.1093/mp/sst038 Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

Figure 1 Functional Analysis of Rice OsOSC6. (A) qRT–PCR relative expression levels of OsOSC6 from rice organs at different developmental stages. Embryo, endosperm, plumule, radicle, coleoptile, seedling root, seedling stem, seedling leaf, mature root, mature stem, mature leaf, leaf node, stamen, pistil, lemma, and palea. OsActin2 was used as a housekeeping gene, relative expression levels were obtained with the ΔΔCt method, and OsOSC6 expression values in embryos were set to a value of 1. Data represent the means ± SD for three biological replicates. (B, C) Pearson correlation analyses of codon usage frequency (CUF) between OsOSC6 (B) and SnOsOSC6 (C), as well as for the genes of P. pastoris (x-axis) from the Kuzusa database. (D, E) Frequencies of the excluded and preferentially included codons in OsOSC6 (D) and SnOsOSC6 (E). (F) Identification of rice α-/β-amyrin synthases in yeast and in planta. GC–MS (TIC, total ion chromatogram; EIC218, extracted ion chromatograms at m/z 218) analyses of extracts of yeast expressing SnOsOSC6 and the empty vector control (pPICZ-A), mature leaves of transgenic rice overexpressing OsOSC6 and wild-type rice, and authentic α- and β-amyrin (reference standards). ‘1’ and ‘2’ represent β-amyrin and α-amyrin, respectively. (G) The mass spectra of trimethylsilyl (TMS) ethers of authentic reference standards of α- and β-amyrin, and the SnOsOSC6 yeast assay product extracts. Molecular Plant 2013 6, 1726-1729DOI: (10.1093/mp/sst038) Copyright © 2013 The Authors. All rights reserved. Terms and Conditions