Transformation Of Resveratrol Synthase Gene In Strawberry

Resveratrol (Res) is a kind of plant phytoalexin usually drived from plant secondary metabolism that has many advantages on human health such as antioxidant, anti-cancer, prevention of cardiovascular diseases and so on. Because of its scarce supply from plant in nature, it is necessary to over-produce this compound in plant by biotechnology. Resveratrol synthase (RS) is the key enzyme in metabolic pathway of Res bio-synthesis. The RS gene under the control of a fruit-specific promoter RJ39 was considered to transform into strawberry as a strategy for improving the nutritional quality of strawberry and the work was reported as below:1. According to the sequence of NCBI, we designed the primers and isolated seven fragments of RS gene with PCR amplification from DNA of seven plants (Ampelopsis aconitifolia, Parthenocissus henryana, P.quinquefolia, P.himalayana, Morus alba L., Humulus scandens and Arachis hypogaea), the results showed that seven PCR products all had 635bp of nucleotide without any introns in their DNA sequences and they encoded 211 amino acids.2. According to the open reading frame(ORF) of RS in grape, we designed the primers and isolated a gene encoding RS from Parthenocissus henryana by RT-PCR method. The full-length cDNA of RS gene was 1179bp, encoding a protein of 392 amino acids.3. We isolated a fruit-specific promoter RJ39 from strawberry by PCR method. The full-length DNA of promoter is 860bp. There are some functional regions including TATA-box, CAAT-box, G-box, I-box and so on.4. Driven by RJ39 promoter and markered by herbicide-resistant gene, the plant binary expression vector for RS gene which was named as pCAMBIA3300-RJ39-RS-Tnos has been built. After introducing the RS gene into strawberry by Agrobacterium-mediated method, the PCR and Southern blotting showed that the RS gene had been transformed into plant.