Abstract
Axenic single cells and honnogon of Spirulina platensis were prepared by treatment with ion mixture of sodium and calcium at the concentration of 500 mmol/1 each, and used as recipients in genetic transformation. In order to inactivate both external and internal nucleases, ethylene diamine tetraacetic acid (EDTA) was supplemented in the medium two days before harvest and filaments were rinsed with the medium upon harvest. Plasmid pBV220 was used as the original vector, and its ampicillin resistance cassette (Amp R) was replaced by chloramphenicol acetyltransferase (Cat) gene from pIJ4813, firefly luciferase gene was placed downstream of the promotors, and polymerase chain reaction amplified fragments from S platensis genome were inserted into its Bgl I site. 4 recombinants were constructed, and pBVCS02L was successfully introduced into S. platensis by electroporation. Electroporated cells were spread on selective plates and algal clones were picked up after cultivation at 25 °C for 30 days. Two clones performed chloramphenicol resistance for 20 generations. The success in genetic transformation of S. platensis was verified by dot blotting and SDS-PAGE.
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Zhang, X.C., Mao, Y.X., Wang, G.G., Zhang, B.H., Yang, G.P., Sui, Z.H. (2001). Preliminary Studies on the Genetic Transformation of Spirulina Platensis . In: Chen, F., Jiang, Y. (eds) Algae and their Biotechnological Potential. Springer, Dordrecht. https://doi.org/10.1007/978-94-015-9835-4_20
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DOI: https://doi.org/10.1007/978-94-015-9835-4_20
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