Expression of Foreign Genes in Dunaliella by Electroporation
Expression of Foreign Genes in Dunaliella by Electroporation
- Molecular Biotechnology, 30, p.185-192, 2005 .
An electroporation procedure has been described for introducing plasmid DNA into Dunaliella salina cells. By this procedure, a bulk of plasmid DNA was delivered into the cells and retained for at least 3 d. Reverse transcriptase polymerase chain reaction (RT-PCR)and sequencing analyses indicated that the transcription and pre-mRNA splicing of ble gene (contributing the Zeocin resistance)were detected in the cells as early as 1 h after the electroporation. Individual colonies could retain the resistance to 10 mg/L Zeocin for at least 6 mo. Subsequent Southern blot analysis showed the existence of introduced plasmid DNA inside these colonies. However, most of the cells (approx 90
DUNALIELLA SALINA
ELECTROPORATION
BLE GENE
ZEOCIN
EPISOMAL DNA
An electroporation procedure has been described for introducing plasmid DNA into Dunaliella salina cells. By this procedure, a bulk of plasmid DNA was delivered into the cells and retained for at least 3 d. Reverse transcriptase polymerase chain reaction (RT-PCR)and sequencing analyses indicated that the transcription and pre-mRNA splicing of ble gene (contributing the Zeocin resistance)were detected in the cells as early as 1 h after the electroporation. Individual colonies could retain the resistance to 10 mg/L Zeocin for at least 6 mo. Subsequent Southern blot analysis showed the existence of introduced plasmid DNA inside these colonies. However, most of the cells (approx 90
DUNALIELLA SALINA
ELECTROPORATION
BLE GENE
ZEOCIN
EPISOMAL DNA