Expression of Foreign Genes in &lt;I&gt;Dunaliella&lt;/I&gt; by Electroporation

Sun, Yu; Yang, Zhiyong; Gao, Xing; Li, Qiyun; Zhang, Qingqi; Xu, Zhengkai

doi:10.1385/mb:30:3:185

Cited by 96 publications

(45 citation statements)

References 13 publications

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“…Members of the chlorophyte group that have been transformed include C. reinhardtii, which has been transformed using a variety of methods (44,87,88,93,170); Chlorella ellipsoidea (22, 83); Chlorella saccharophila (108); C. vulgaris (30, 69); Haematococcus pluvialis (177,187); V. carteri (81,163); Chlorella sorokiniana (30); Chlorella kessleri (49); Ulva lactuca (76); Dunaliella viridis (180); and D. salina (181,183). Heterokontophytes that have reportedly been transformed include Nannochloropsis oculata (21); diatoms such as T. pseudonana (147), P. tricornutum (2,210,211), Navicula saprophila (45), Cylindrotheca fusiformis (52,148), Cyclotella cryptica (45), and Thalassiosira weissflogii (51); and phaeophytes, such as Laminaria japonica (150) and Undaria pinnatifada (151).…”

Section: Genetic Engineering Of Microalgaementioning

confidence: 99%

Genetic Engineering of Algae for Enhanced Biofuel Production

et al. 2010

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2There are currently intensive global research efforts aimed at increasing and modifying the accumulation of lipids, alcohols, hydrocarbons, polysaccharides, and other energy storage compounds in photosynthetic organisms, yeast, and bacteria through genetic engineering. Many improvements have been realized, including increased lipid and carbohydrate production, improved H 2 yields, and the diversion of central metabolic intermediates into fungible biofuels. Photosynthetic microorganisms are attracting considerable interest within these efforts due to their relatively high photosynthetic conversion efficiencies, diverse metabolic capabilities, superior growth rates, and ability to store or secrete energy-rich hydrocarbons. Relative to cyanobacteria, eukaryotic microalgae possess several unique metabolic attributes of relevance to biofuel production, including the accumulation of significant quantities of triacylglycerol; the synthesis of storage starch (amylopectin and amylose), which is similar to that found in higher plants; and the ability to efficiently couple photosynthetic electron transport to H 2 production. Although the application of genetic engineering to improve energy production phenotypes in eukaryotic microalgae is in its infancy, significant advances in the development of genetic manipulation tools have recently been achieved with microalgal model systems and are being used to manipulate central carbon metabolism in these organisms. It is likely that many of these advances can be extended to industrially relevant organisms. This review is focused on potential avenues of genetic engineering that may be undertaken in order to improve microalgae as a biofuel platform for the production of biohydrogen, starch-derived alcohols, diesel fuel surrogates, and/or alkanes.

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Section: Genetic Engineering Of Microalgaementioning

confidence: 99%

Genetic Engineering of Algae for Enhanced Biofuel Production

et al. 2010

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“…This technique involves bombardment with DNA-coated microprojectiles and has been successfully used for a variety of algae including green algae and diatoms and is also the method of choice for chloroplast or mitochondrial genome transformation (Apt et al 1996;Remacle et al 2006;Kroth 2007). Other methods to create transgenic algae are the agitation of cells in the presence of glass beads and DNA, agitation with silicon-carbide whiskers and electroporation (Kindle 1990;Dunahay et al 1997;Sun et al 2005). More recent developments for the improved overexpression of transgenes involve the use of vectors containing nuclear matrix attachment regions to increase the expression level of foreign genes which has been carried out in the halotolerant alga D. salina .…”

Section: Genetic Engineering To Improve Algal Productivitymentioning

confidence: 99%

Microalgae as second generation biofuel. A review

Singh¹,

Dhar²

2011

Agron. Sustain. Dev.

155

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Microalgae are autotrophic microorganisms having extremely high photosynthetic efficiency and are valued as rich source of lipids, hydrocarbons, and other complex oils for biodiesel besides being an invaluable source of bioethanol, biomethane, and biohydrogen. Biodiesel produced from oilseed crops such as jatropha and soy have lower yields per unit land area and threaten food security. Indeed, microalgae have higher oil yields amounting to about 40 times more oil per unit area of land in comparison to terrestrial oilseed crops such as soy and canola. Further, microalgae production does not require arable land for cultivation. Biofuel is regarded as a proven clean energy source and several entrepreneurs are attempting to commercialize this renewable source. Technology for producing and using biofuel has been known for several years and is frequently modified and upgraded. In view of this, a review is presented on microalgae as second generation biofuel. Microalgal farming for biomass production is the biggest challenge and opportunity for the biofuel industry. These are considered to be more efficient in converting solar energy into chemical energy and are amongst the most efficient photosynthetic plants on earth. Microalgae have simple cellular structure, a lipid-rich composition, and a rapid rate of reproduction. Many microalgal strains can be grown in saltwater and other harsh conditions. Some autotrophic microalgae can also be converted to heterotrophic ones to accumulate high quality oils using organic carbon. However, there are several technical challenges that need to be addressed to make microalgal biofuel profitable. The efficiency of microalgal biomass production is highly influenced by environmental conditions, e.g., light of proper intensity and wavelength, temperature, CO 2 concentration, nutrient composition, salinities and mixing conditions, and by the choice of cultivation systems: open versus closed pond systems, photobioreactors. Currently, microalgae for commercial purpose are grown mostly in open circular/elongated "raceway" ponds which generally have low yields and high production costs. However, a hybrid system combining closed photobioreactor and open pond is a better option. The biggest hurdle in commercialization of microalgal biofuel is the high cost and energy requirement for the microalgal biomass production, particularly agitation, harvesting, and drying of biomass. In order to conserve energy and reduce costs, algae are often harvested in a two-step process involving flocculation followed by centrifugation, filtration, or micro-straining to get a solid concentration. However, the major bottlenecks in algal biodiesel production within the cell can be identified and handled by adopting a system approach involving transcriptomics, proteomics, and metabolomics. Research and developments in the field of new materials and advanced designs for cultivation in closed bioreactors, use of waste water for biomass production, screening of efficient strains, high-value coproduct strategy,...

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“…A population of D. salina CCAP 19/18 cells was harvested from a 250-ml culture in its exponential growth phase (approximately 1 × 10 6 cells ml -1 ), washed repeatedly with pre-electroporation buffer (0.2 M mannitol, 0.2 M sorbitol) to remove residual salts, and resuspended in electroporation buffer (0.08 M KCl, 0.005 M CaCl 2 , 0.01 M HEPES, 0.2 M mannitol, 0.2 M sorbitol) to achieve a final density of 8 × 10 7 cells ml -1 (Sun et al, 2005). Next, 500 l electroporation samples were prepared in 0.4 cm electrode gap cuvettes (Bio-Rad), which contained either 4 × 10 7 or 1 × 10 6 cells ml -1 , 20 mg ml -1 of pSP124, and 2 mg ml -1 of herring or salmon sperm DNA (Sigma).…”

Section: Electroporationmentioning

confidence: 99%