[Transformation of Chlamydomonas reinhardtii CW-15 with the hygromycin phosphotransferase gene as a selective marker].
To transform Chlamydomonas reinhardtii Dang. Cells, plasmid pCTVHyg was constructed with the use of the Escherichia coli hygromycin phosphotransferase gene (hpt) controlled by the SV40 early promoter. Cells of the CW-15 mutant strain were transformed by electroporation, with the yield reaching 10(3) hygromycin-resistant (HygR) clones per 10(6) recipient cells. The exogenous DNA integrated in the Ch. reinhardtii nuclear genome showed stable transmission for approximately 350 cell generations, while hygromycin resistance was expressed as an unstable character. Codon usage was compared for the hpt gene and Ch. reinhardtii nuclear genes. The results testified that codon usage bias, which is characteristic of Ch. reinhardtii, is not the major factor affecting foreign gene expression. The advantages of the selective system for studying Ch. reinhardtii transformation with heterologous genes are discussed.
Ladygin
V G
VG
Institute of the Fundamental Problems of Biology, Russian Academy of Sciences, Pushchino, Moscow Oblast, 142290 Russia. ladygin@issp.serpukhov.su
Butanaev
A M
AM
D000818:Animals; D016825:Chlamydomonas reinhardtii / Q000235:genetics*; D003062:Codon; D018274:Electroporation; D005819:Genetic Markers; D009154:Mutation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics*; D014170:Transformation, Genetic
D003062:Codon; D005819:Genetic Markers; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase