[The transformation of the unicellular alga Chlamydomonas reinhardtii by electroporation].
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 Tn5 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) HygR 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.
Ladygin
V G
VG
Institute of Basic Biological Problems, Russian Academy of Sciences, Pushchino, Moscow Oblast, 142290 Russia. ladygin@issp.serpukhov.su
D000818:Animals; D016825:Chlamydomonas reinhardtii / Q000235:genetics*; D004251:DNA Transposable Elements; D018274:Electroporation; D004926:Escherichia coli / Q000235:genetics; D009154:Mutation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D013539:Simian virus 40 / Q000235:genetics; D014170:Transformation, Genetic
D004251:DNA Transposable Elements; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase