Advanced multigene expression system for Nannochloropsis salina using 2A self-cleaving peptides

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Even though there has been much interest in genetic engineering of microalgae, its progress has been slow due to the difficulty and limitation of available techniques. Currently, genetic modification in most microalgal strains is confined to single gene transformation. Here, a multigene expression system for the oleaginous model strain Nannochloropsis salina was developed with glycine-serine-glycine spacer linked 2A self-cleaving peptides (2A) for the first time. An efficiency test of the four most widely used 2As revealed that two different types of 2As T2A and E2A have the best performance in N. salina with a maximum cleavage rate of nearly 45%. The system was able to express the linked sequence of the selection marker shble and the fluorescence protein sfCherry with intact functions. Because 2A enabled multigene expression in the single cassette form, the use of 2A also reduced the vector size, which along with the stronger promoter resulted in a 9-fold increase in the transformation efficiency. Furthermore, confirmative screening accuracy of more than 90% was observed. Hence, the 2A applied vector system is expected to be beneficial in microalgal research field because it enables multigene expression as well as offering improved transformation and screening efficiency.
Publisher
ELSEVIER SCIENCE BV
Issue Date
2018-07
Language
English
Article Type
Article
Citation

JOURNAL OF BIOTECHNOLOGY, v.278, pp.39 - 47

ISSN
0168-1656
DOI
10.1016/j.jbiotec.2018.04.017
URI
http://hdl.handle.net/10203/242594
Appears in Collection
CBE-Journal Papers(저널논문)
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