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High-efficiency nuclear transformation of the oleaginous marine Nannochloropsis species using PCR product
Li, Fengjuan1; Gao, Dawen1; Hu, Hanhua2
2014-05-01
Source PublicationBIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
ISSN0916-8451
Volume78Issue:5Pages:812-817
AbstractNannochloropsis 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.
SubtypeArticle
KeywordElectroporation Nannochloropsis Transformation Cotransformation Gus
DOI10.1080/09168451.2014.905184
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine ; Physical Sciences
Indexed BySCI
Language英语
WOS Research AreaBiochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Chemistry ; Food Science & Technology
WOS SubjectBiochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Chemistry, Applied ; Food Science & Technology
WOS IDWOS:000339070600014
WOS KeywordDIATOM PHAEODACTYLUM-TRICORNUTUM ; FATTY-ACID-COMPOSITION ; MICROALGAE ; GROWTH ; EUSTIGMATOPHYCEAE ; TRIACYLGLYCEROLS ; CHLAMYDOMONAS ; AQUACULTURE ; GENES
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Cited Times:30[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/20406
Collection藻类生物学及应用研究中心_期刊论文
Affiliation1.Harbin Inst Technol, State Key Lab Urban Water Resource & Environm, Harbin 150006, Peoples R China
2.Chinese Acad Sci, Inst Hydrobiol, Key Lab Algal Biol, Wuhan, Peoples R China
Recommended Citation
GB/T 7714
Li, Fengjuan,Gao, Dawen,Hu, Hanhua. High-efficiency nuclear transformation of the oleaginous marine Nannochloropsis species using PCR product[J]. BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY,2014,78(5):812-817.
APA Li, Fengjuan,Gao, Dawen,&Hu, Hanhua.(2014).High-efficiency nuclear transformation of the oleaginous marine Nannochloropsis species using PCR product.BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY,78(5),812-817.
MLA Li, Fengjuan,et al."High-efficiency nuclear transformation of the oleaginous marine Nannochloropsis species using PCR product".BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 78.5(2014):812-817.
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