Expression of foreign genes in Dunaliella by electroporation.
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.
Institute of Plant Physiology & Ecology, Shanghai Institute for Biological Sciences, The Chinese Academy of Sciences, Shanghai.
Yu
Y
Sun
Zhiyong
Z
Yang
Xiaoshu
X
Gao
Qiyun
Q
Li
Qingqi
Q
Zhang
Zhengkai
Z
Xu
D001761:Bleomycin / Q000494:pharmacology; D015139:Blotting, Southern; D002470:Cell Survival / Q000235:genetics; D000460:Chlorophyta / Q000235:genetics* / Q000378:metabolism*; D004247:DNA / Q000235:genetics; D004351:Drug Resistance / Q000187:drug effects; D018274:Electroporation / Q000379:methods*; D015870:Gene Expression / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D014170:Transformation, Genetic
D012333:RNA, Messenger; D001761:Bleomycin; C105427:Zeocin; D004247:DNA