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N-terminal sequences affect expression of triterpene biosynthesis enzymes in Chlamydomonas chloroplasts

Algal Research. 2019; 
Hsu Shih-Chi, Browne Daniel R, Tatli Mehmet, Devarenne Timothy P, Stern David B
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Catalog Antibody After centrifugation, cell pellets were resuspended in protein sample buffer and heated to 100?°C for 7?min, and 1?×?106?cells were analyzed by SDS-PAGE and immunoblotting. Various epitope antibodies were used to detect tagged transgenic proteins: anti-c-Myc 9E10 antibody (626801; BioLegend, San Diego, CA), THE? His-tag antibody (A00186; GenScript, Piscataway, NJ), anti-FLAG M2 antibody (F1804; Sigma), anti-FLAG polyclonal antibody (F7425; Sigma), and THE? HA-tag antibody (A01244; Genscript). Get A Quote

摘要

Metabolic engineering is an emerging technology to modify the biochemical properties of living cells. In microalgae, metabolic engineering has often been directed towards optimizing the production of desirable lipids or related bioproducts. Here we describe efforts to engineer the green alga?Chlamydomonas reinhardtii?for the production of botryococcene, a drop-in biofuel precursor. Genes encoding farnesyl diphosphate synthase (FPS) and squalene synthase-like (SSL)-1 and -3, were introduced into the chloroplast genome using biolistic transformation. Through a series of construct modifications, we identified intergenic sequences that promote expression of stable, discrete transcripts. We also found amino acids ... More

關(guān)鍵詞

ChloroplastProtein expressionN-terminal sequenceAlgaeChlamydomonas reinhardtiiBotryococcene
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