Abstract
Synechococcus sp. CC9311 is a marine cyanobacterium characterized by type IV chromatic acclimation (CA). A genetic transformation system was developed as a first step to elucidate the molecular mechanism of CA. The results show that Synechococcus sp. CC9311 cells were sensitive to four commonly used antibiotics: ampicillin, kanamycin, spectinomycin, and chloramphenicol. An integrative plasmid to disrupt the putative phycoerythrin lyase gene mpeV, using a kanamycin resistance gene as selectable marker, was constructed by recombinant polymerase chain reaction. The plasmid was then transformed into Synechococcus sp. CC9311 via electroporation. High transformation efficiency was achieved at a field strength of 2 kV/cm. DNA analysis showed that mpeV was fully disrupted following challenge of the transformants with a high concentration of kanamycin. In addition, the transformants that displayed poor growth on agar SN medium could be successfully plated on agarose SN medium.
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Supported by the Key Innovation Project of Institute of Oceanology, Chinese Academy of Sciences (No. 2009-2), the Natural Science Foundation of Shandong Province (No. 2009ZRB02542), the Foundation of Key Laboratory of Marine Bioactive Substance and Modern Analytical Techniques, SOA (No. MBSMAT-2010-03), the National Natural Science Foundation of China (No. 41276164), and the Natural Science Foundation of Jiangsu Province (No. BK2012650)
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Chen, H., Lin, H., Jiang, P. et al. Genetic transformation of marine cyanobacterium Synechococcus sp. CC9311 (Cyanophyceae) by electroporation. Chin. J. Ocean. Limnol. 31, 416–420 (2013). https://doi.org/10.1007/s00343-013-2164-5
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DOI: https://doi.org/10.1007/s00343-013-2164-5