Investigation on Electroporation: Mediated Transformation of Chlorella ellipsoidea
Chlorella ellipsoidea is a kind of micro algae biological reactor. It is a suitable eukaryotic expression system for foreign proteins, and the bioactivity of the expressed protein is similar to the natural counterpart. However, introduction of foreign gen
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Investigation on Electroporation: Mediated Transformation of Chlorella ellipsoidea Zhihan Zuo, Feifei Qin, Yichen Zhang, Yichen Liu and Jinsheng Sun
Abstract Chlorella ellipsoidea is a kind of micro algae biological reactor. It is a suitable eukaryotic expression system for foreign proteins, and the bioactivity of the expressed protein is similar to the natural counterpart. However, introduction of foreign gene into C. ellipsoidea cells is the precondition for its efficient expression. In this paper the electroporation conditions to transform C. ellipsoidea including cultured medium, growth rate of C. ellipsoidea, voltage strength were systematically investigated. BG11 without Mg2+ was selected as the medium for cultivating C. ellipsoidea. The exponential phase cells were harvested and electroporated under different electric voltages. Voltage at the 630 V was identified to be optimal by determining the cells viability after electroporation. Under this condition, the plasmid pSC2 carrying gfp was successfully transformed into C. ellipsoidea. The gfp gene could continuously express for at least 7 days. (This work was financially supported by grants from the National Basic Research Development Program of China (973 programs, 2012CB114405), National HighTech Research and Development Program of China (863 programs, 2012AA092205 and 2012AA10A401), National Key Technology Research and Development Program (2011BAD13B07 and 2011BAD13B04), Doctoral found of Tianjin normal university: 52X09010 and Open research found for city stage key laboratory of Tianjin normal university). Keywords Cell viability Voltage strength
Chlorella ellipsoidea Electroporation Medium
Z. Zuo F. Qin Y. Zhang Y. Liu J. Sun (&) College of Life Science, Tianjin Normal University, Tianjin Key Laboratory of Animal and Plant Resistance, Tianjin 300387, People’s Republic of China e-mail: [email protected]
T.-C. Zhang et al. (eds.), Proceedings of the 2012 International Conference on Applied Biotechnology (ICAB 2012), Lecture Notes in Electrical Engineering 249, DOI: 10.1007/978-3-642-37916-1_28, Ó Springer-Verlag Berlin Heidelberg 2014
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28.1 Introduction Unicellular green algea Chlorella ellipsoidea has long been regarded as a potential diet source due to its high protein content and various physiologically active substances [1]. This species is an essential phytoplankton used as live feed for fisheries with a high content of proteins and fatty acids. In addition, due to its fast growth and low cost, C. ellipsoidea is thus a promising candidate bioreactor for the large-scale production of value-added proteins. To make a Chlorella microalgal bioreactor, an appropriate genetic transformation system is needed to ensure the expression of the target gene in transgenic Chlorella. In the case of Chlorella, development of genetic transformation has been slow. Many techniques are available for gene transformation of plant cells. Such as protoplasting, electroporation, particle bombardment, and Agrobacterium-mediated
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