Expression of foreign genes in dunaliella by electroporation

• Published in: Molecular biotechnology 2005-07, Vol.30 (3), p.185-192
• Main Authors: Sun, Yu, Yang, Zhiyong, Gao, Xiaoshu, Li, Qiyun, Zhang, Qingqi, Xu, Zhengkai
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Biotechnology; Bleomycin - pharmacology; Blotting, Southern; Cell Survival - genetics; Chlorophyta - genetics; Chlorophyta - metabolism; Deoxyribonucleic acid; DNA; DNA - genetics; Drug Resistance - drug effects; Dunaliella; Dunaliella salina; Electroporation - methods; Fundamental and applied biological sciences. Psychology; Gene expression; Gene Expression - genetics; Microbiology; Plasmids; Plasmids - genetics; RNA, Messenger - genetics; Studies; Transformation, Genetic
• ISSN: 1073-6085; 1559-0305; 1073-6085
• DOI: 10.1385/mb:30:3:185

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%) lost the resistance in the presence of 5 mg/L Zeocin during subculturing, which was consistent with the observations of both rearranged and episomal plasmid DNA existed in the cells. Nevertheless, the electroporation procedure allows introducing a gene of interest and studying its expression and function in D. salina cells.