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Title Optimum Conditions for Transformation of Synechocystis sp. PCC 6803
Author Xiaonan Zang, Bin Liu, Shunmei Liu, K.K.I.U. Arunakumara, and Xuecheng Zhang*
Address College of Marine Life Sciences, Ocean University of China, Qingdao 266003, P. R. China
Bibliography Journal of Microbiology, 45(3),241-245, 2007,
Key Words Cyanobacterium, electroporation, Synechocystis sp. PCC 6803, ultrasonic
Abstract This study was conducted to determine the optimal conditions for introduction of exogenous DNA into Synechocystis sp. PCC 6803. Of the three transformation techniques studied, electroporation, ultrasonic transformation and natural transformation, natural transformation showed the highest efficiency. Additionally, this study demonstrated that the higher plasmid concentration and longer homologous recombining fragments resulted in a greater number of transformants. For successful transformation, the lowest concentration of plasmid was 0.02 μg/ml, and the shortest homologous recombining fragment was 0.2 kb. Use of Synechocystis sp. PCC 6803 in the logarithmic growth phase resulted in two-fold higher transformation rate than that of the same organism when cells in the latent phase or the plateau phase were used for transformation. Pretreatment of the host strain, Synechocystis sp. PCC 6803, with EDTA (2 mM) for two days prior to transformation increased the transformation efficiency by 23%. Additionally, incubation of the cells and DNA for 5 h under light conditions increased the transformation efficiency by two orders of magnitude. Moreover, recovery treatment of the cells before they were plated onto antibiotic medium also increased the transformation efficiency.
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