Journal
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
Volume 78, Issue 5, Pages 812-817Publisher
TAYLOR & FRANCIS LTD
DOI: 10.1080/09168451.2014.905184
Keywords
electroporation; Nannochloropsis transformation; cotransformation; GUS
Categories
Funding
- National Natural Science Foundation of China [40976079]
- National Key Basic Research Project of China [2011CB 200901]
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Nannochloropsis are model species for investigating biofuel production by algae. To develop them into an integrated photons-to-fuel production platform, high efficiency transformation methods are necessary. Here, we obtained the beta-tubulin promoter regions of all recognized species of genus Nannochloropsis, and successfully transformed all five marine species by electroporation. In addition, the PCR amplified double stranded DNA fragments (PCR fragments) based transformation system was established in these Nannochloropsis species, which showed much higher transformation efficiency (10.7-61.2 x 10 (6), 1.5-13-fold) than that of linearized plasmid based transformation. The cotransformation of N. salina using a circular plasmid containing a non-selectable GUS gene and a PCR fragment containing only a selection marker cassette was also achieved and found to be very efficient (over 50%). This simple and highly efficient transformation protocol reported in our study provided a useful tool for gene functional analysis and genetic engineering of the oleaginous Nannochloropsis species.
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