Towards a synthetic nuclear transcription system in green algae: Characterization of Chlamydomonas reinhardtii nuclear transcription factors and identification of targeted promoters

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• 作者：Melissa S. Anderson ; Travis J. Muff ; David R. Georgianna ; Stephen P. Mayfield ; ^{smayfield@ucsd.edu}
• 关键词：Chlamydomonas ; Transcription factor ; Nuclear gene expression ; RNA-sequencing ; Yeast one-hybrid ; Synthetic biology
• 刊名：Algal Research
• 出版年：2017
• 出版时间：March 2017
• 年：2017
• 卷：22
• 期：Complete
• 页码：47-55
• 全文大小：1056 K
• 卷排序：22

文摘

Algae represent a unique opportunity to develop a sustainable production platform for many valuable products. A key hindrance to exploiting algae's potential as a bio-product manufacturing platform is our limited understanding of nuclear genetic regulatory systems. To begin to address this deficiency we identified prospective transcription factor-promoter pairs that have the potential for use in a controllable synthetic transcription system. We constructed a library of 92 recombinant nuclear transcription factors (TFs) from the model alga Chlamydomonas reinhardtii. Using a yeast one-hybrid assay, we identified TFs with the ability to activate transcription and chose TF64 for further analysis. We generated two independent C. reinhardtii strains that constitutively expressed the gene encoding TF64 and used RNA-sequencing on these strains to identify transcripts that were altered by expression of the TF's gene. Many mRNAs were regulated by TF64, and we confirmed activation of transcription of nine genes from the LHCBM family by RT-qPCR. Overall, we were able to identify numerous potential TF-cognate promoter binding partners in C. reinhardtii, thus laying the groundwork to develop a controllable synthetic nuclear transcription system for microalgae, an essential tool for realizing their full biotechnological potential.