Journal
MICROBIOLOGY
Volume 72, Issue 5, Pages 585-591Publisher
MAIK NAUKA/INTERPERIODICA
DOI: 10.1023/A:1026051402366
Keywords
plasmid pCTVHyg; hytgromycin phosphotransferase; transformation; electroporation; mutant CW-15; Chlamydomonas reinhardtii
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The cell wall-lacking mutant CW-15 of the unicellular green alga Chlamydomonas reinhardtii was transformed by electroporation using plasmid pCTVHyg, which was constructed with the hygromycin phosphotransferase gene hpt as the selective marker and the Tn(5) transposon of Escherichia coli under the control of the virus SV40 early gene promoter. Under optimal conditions (10(6) mid-exponential cells/ml; electric field strength 1 kV/cm; and pulse length 2 ms), the transformation yielded 10(3) Hyg(R) transformants per 10(6) recipient cells. The exogenous DNA integrated into the nuclear genome of Ch. reinhardtii was persistently inherited through more than 350 cell generations. The advantages of this system for the transformation of Ch. reinhardtii with heterologous genes are discussed.
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