[An efficient way for obtaining transformants of Chlamydomonas reinhardtii by electroporation].
The cells of mutant CW-15, which does not form the cell wall, were transformed by electroporation. It was found that the transformation was optimal at a suspension density of 10(6) cells/ml in the middle of the logarithmic growth phase at a field intensity of 1 kW/cm and pulse duration 2 ms. It was shown that, under these conditions, the efficiency of transformation reached 10(3) HygR-clones per 10(6) recipient cells. Exogenic DNA integrated into the genome of the nucleus of Ch. reinhardtii was inherited constantly for more than 350 generations; however, the tolerance to hygromycin appeared as an unstable feature. The advantages of using mutant CW-15 and the selective system to study the transformation of Ch. reinhardtii by heterologous genes are discussed.
Ladygin
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D000818:Animals; D016825:Chlamydomonas reinhardtii / Q000235:genetics*; D018274:Electroporation; D004926:Escherichia coli; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D014170:Transformation, Genetic
D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase