Journal
BIOTECHNOLOGY JOURNAL
Volume 11, Issue 3, Pages 384-392Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/biot.201500269
Keywords
Chlamydomonas transformation; Dunaliella LIP promoter; GT-1 binding motif; Light-inducible motif; Sequence over-represented in light-induced promoter (SORLIP)
Funding
- Korea CCS RD Center (KCRC) [NRF-2014M1A8A1049273]
- Korean Government (Ministry of Science, Ict. & Future Planning) [NRF-2013R1A2A1A01015644]
- Energy Efficiency & Resources Core Technology Program of Korea Institute of Energy Technology Evaluation and Planning (KETEP)
- Ministry of Trade, Industry & Energy, Republic of Korea [20142020200980]
- Korea Evaluation Institute of Industrial Technology (KEIT) [20142020200980] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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Promoter of the light-inducible protein gene (LIP) of Dunaliella was recently isolated in our laboratory. The aim of this work is to find the light-inducible motif in the Dunaliella LIP promoter and verify its regulatory motif with a Gaussia luciferase reporter gene transformed in Chlamydomonas reinhardtii. 400 bp upstream to the translational start site of the Dunaliella LIP gene was gradually truncated and analyzed for the luciferase expression. Furthermore, this promoter comprising duplicated or triplicated light-responsive motifs was tested for its augmentation of light response. Two putative light-responsive motifs, GT-1 binding motif and sequences over-represented in light-repressed promoters (SORLIP) located in the 200 bp LIP promoter fragment were analyzed for their light responsibility. It is turned out that SORLIP was responsible for the light-inducible activity. With the copy number of SORLIP up to three showed stronger high light response compared with the native LIP promoter fragment. Therefore, we found a light-responsive DNA motif operating in Chlamydomonas and confirm a synthetic promoter including this motif displayed light inducibility in heterologously transformed green algae for the first time. This light-inducible expression system will be applied to various area of algal research including algal biotechnology.
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