|Inhibition of pds Gene Expression via the RNA Interference Approach in Dunaliella salina (Chlorophyta)|
Sun; Zhang; Sui; Mao (2008). Inhibition of pds Gene Expression via the RNA Interference Approach in Dunaliella salina (Chlorophyta). Mar. Biotechnol. 10(3): 219-226
In: Marine Biotechnology. Springer-Verlag: New York. ISSN 1436-2228, more
Gene expression; Dunaliella salina (Dunal) Teodoresco, 1905 [WoRMS]; Marine
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To investigate the potential of double-stranded RNA interferencing with gene expression in Dunaliella salina, a plasmid pBIRNAI-Dsa was constructed to express hairpin RNA (hpRNA) containing sequences homologous to phytoene desaturase gene (pds), a key gene in carotenoid biosynthesis, and transformed into D. salinaby electroporation. The relative transcription level of pds in pBIRNAI-Dsa-treated cells to nontreated cells was quantitated and the gene silencing efficiency by RNAi was evaluated via real-time polymerase chain reaction (PCR). The transcriptions of pds of the pBIRNAI-Dsa-treated group changed compared to those of the control group, and the was lowest on the 7th day, corresponding to 0.281265-fold of the relative pds control transcript; a relatively strong gene inhibition effect was therefore deduced. The transcript of pds may be modulated in a wide range, and a reduced transcription even to 28% of the normal level may be tolerated for its survival. This study shows that dsRNA-mediated genetic interference can induce sequence-specific inhibition of gene expression and pBIRNAI-Dsa can be used for transient suppression of gene expression inD. salina. The aim of this study was to exploit dsRNA-mediated gene silencing and to provide a foundation for gene function research in D. salina.