Efficiency of different heterologous promoters in the unicellular microalga Chlamydomonas reinhardtii
- 14 January 2013
- journal article
- research article
- Published by Wiley in Biotechnology Progress
- Vol. 29 (2), 319-328
- https://doi.org/10.1002/btpr.1690
Abstract
Despite the biotechnological interest of microalgae, no robust and stable methods for genetic transformation of most microalgal strains exist. The scanty and disperse data about the efficiency of heterologous promoters in microalgae and the use of different transformation methods, DNA quantities and reporter genes in the existing studies makes very difficult a real comparison of their efficiency. Using Chlamydomonas reinhardtii as a host, we have evaluated the efficiency of the heterologous promoters of cauliflower mosaic virus 35S (CaMV 35S) and Agrobacterium nopaline synthase (NOS) genes. These promoters were fused to the paromomycin conferring‐resistance aminoglycoside 3′‐phosphotransferase encoding gene (APHVIII), and C. reinhardtii was transformed by the glass beads agitation method. The transformation efficiency and the APHVIII transcript and protein levels were evaluated in a series of transformants for each promoter. The chimeric promoter HSP70A/RBCS2 and the promoter‐less APHVIII marker gene were used for comparison. We found significantly higher transformation efficiencies and higher level of APHVIII expression in those transformants harboring the NOS promoter than in those transformed with CaMV 35S promoter. The NOS promoter, widely used for genetic manipulation of higher plants, has been very rarely used for the transformation of microalgae. The results shown here suggest the possibilities of this heterologous promoter as an efficient system for the genetic manipulation of microalgae. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29: 319–328, 2013Keywords
This publication has 60 references indexed in Scilit:
- Draft genome sequence and genetic transformation of the oleaginous alga Nannochloropsis gaditanaNature Communications, 2012
- High-efficiency homologous recombination in the oil-producing alga Nannochloropsis sp.Proceedings of the National Academy of Sciences, 2011
- Genetic Engineering of Algae for Enhanced Biofuel ProductionEukaryotic Cell, 2010
- Clocks in the Green Lineage: Comparative Functional Analysis of the Circadian Architecture of the Picoeukaryote OstreococcusPlant Cell, 2009
- Towards genetic improvement of commercially important microalga Haematococcus pluvialis for biotech applicationsJournal of Applied Phycology, 2009
- Gaussia-luciferase as a sensitive reporter gene for monitoring promoter activity in the nucleus of the green alga Chlamydomonas reinhardtiiMolecular Genetics and Genomics, 2008
- Transformation of the Green Alga Haematococcus pluvialis with a Phytoene Desaturase for Accelerated Astaxanthin BiosynthesisApplied and Environmental Microbiology, 2006
- Swapped green algal promoters: aphVIII-based gene constructs with Chlamydomonas flanking sequences work as dominant selectable markers in Volvox and vice versaPlant Cell Reports, 2006
- A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindingAnalytical Biochemistry, 1976
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970