TY - JOUR
T1 - Establishment of an efficient genetic transformation system in Scenedesmus obliquus
AU - Guo, Suo Lian
AU - Zhao, Xin Qing
AU - Tang, Ying
AU - Wan, Chun
AU - Alam, Md Asraful
AU - Ho, Shih Hsin
AU - Bai, Feng Wu
AU - Chang, Jo Shu
N1 - Funding Information:
This work was supported by financial support from National Basic Research Program of China ( 2011CB200905 ) as well as the support from Taiwan's Ministry of Economic Affairs (Grant No. 101-D0204-3 ) and Taiwan's National Science Council (Grant No. NSC 101-3113-E-006-015 and NSC 101-3113-E-006-016 ). The authors acknowledge Dr. Qiao Su in Dalian University of Technology, China, for providing the plasmid of pCAMBIA 1302.
PY - 2013/1/10
Y1 - 2013/1/10
N2 - Scenedesmus obliquus belongs to green microalgae, which is attracting attention as a feedstock for biofuels production and biorefinery as well as in bioremediation of environmental pollutants, making its genetic modifications for more efficient growth and accumulation of aimed metabolites significant. However, the genetic transformation system of S. obliquus is still not well established. In the current work, S. obliquus was transformed via electroporation using a plasmid containing chloramphenicol resistance gene (CAT) as a selectable marker and the green fluorescent protein gene (gfp) as a reporter. Using the optimized transformation conditions, the transformation efficiency was 494±48 positive transgenic clones per 106 recipient cells, which is more efficient comparing with those reported in other microalgal transformation studies. Green fluorescence was observed after six months of cultivation, and CAT-specific products were also detected in the transformants by PCR, Southern blot and RT-PCR analysis. This is the first report on establishing such an efficient and stable transformation system for S. obliquus, a prerequisite for both functional genomic studies and strain improvement for other biotechnology applications of this important microalgal species.
AB - Scenedesmus obliquus belongs to green microalgae, which is attracting attention as a feedstock for biofuels production and biorefinery as well as in bioremediation of environmental pollutants, making its genetic modifications for more efficient growth and accumulation of aimed metabolites significant. However, the genetic transformation system of S. obliquus is still not well established. In the current work, S. obliquus was transformed via electroporation using a plasmid containing chloramphenicol resistance gene (CAT) as a selectable marker and the green fluorescent protein gene (gfp) as a reporter. Using the optimized transformation conditions, the transformation efficiency was 494±48 positive transgenic clones per 106 recipient cells, which is more efficient comparing with those reported in other microalgal transformation studies. Green fluorescence was observed after six months of cultivation, and CAT-specific products were also detected in the transformants by PCR, Southern blot and RT-PCR analysis. This is the first report on establishing such an efficient and stable transformation system for S. obliquus, a prerequisite for both functional genomic studies and strain improvement for other biotechnology applications of this important microalgal species.
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U2 - 10.1016/j.jbiotec.2012.10.020
DO - 10.1016/j.jbiotec.2012.10.020
M3 - Article
C2 - 23147423
AN - SCOPUS:84870376406
SN - 0168-1656
VL - 163
SP - 61
EP - 68
JO - Journal of Biotechnology
JF - Journal of Biotechnology
IS - 1
ER -