[Use of the hygromycin phosphotransferase gene as the dominant selective marker for Chlamydomonas reinhardtii transformation].
The hygromycin phosphotransferase gene (hpt) from E. coli under the control of the SV40 early promoter was used as a dominant selectable marker for transformation of Chlamydomonas reinhardtii. Cells were transformed by electroporation (pulse length, 2 ms, field strength, 1 kV/cm). The culture growth phase was a crucial parameter for transformation (optimal density approximately 10(6) cells/ml). It was possible to obtain approximately 10(3) Hyg-resistant colonies under these conditions. Foreign DNA integrated into the Chlamydomonas genome was maintained for at least 8 months but the Hyg-resistant phenotype of the transformed clones was unstable. The frequency of codon usage in the hpt gene was compared with the one in Chlamydomonas nuclear genes. It is supposed that highly biased codon usage in Chlamydomonas does not preclude expression. Advantages of this selection system for studying Chlamydomonas transformation by heterologous genes are discussed.
Butanaev
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AM
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D016825:Chlamydomonas reinhardtii / Q000235:genetics*; D003001:Cloning, Molecular; D003062:Codon; D018274:Electroporation; D004926:Escherichia coli / Q000235:genetics; D005799:Genes, Dominant; D005819:Genetic Markers; D010641:Phenotype; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics*; D011401:Promoter Regions, Genetic; D013539:Simian virus 40 / Q000235:genetics; D014170:Transformation, Genetic
D003062:Codon; D005819:Genetic Markers; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase
Ispol'zovanie gena gigromitsinfosfotransferazy v kachestve dominantnogo selektivnogo markera dlia transformatsii Chlamydomonas reinhardtii.