Transformation of pichia pastoris with resveratrol synthase gene cloning from grape

Aim To construct the Pichia pastoris expression system of RS gene. Methods By PCR,enzyme digestion, ligation and electroporation, construct recombinant vector and transform Pichia pastoris GS115. Results The target fragment was amplified from the grape genome DNA and cloned into pBS-T Vector. ; The recombinant vector pPIC3. 5K/RS was successfully constructed; The target gene was integrated into Pichia pastoris genome via homologous recombination . The results detected by sequencing and PCR show that the Pichia pastoris expression system of RS gene was successfully constructed. Conclusion The obtained method didnot reed high quality RNA, which lowered the experinment condition and made the process quick and simple.