Objective:To construct the plant expression vector including Adketo gene from Adonis aestivalis and E8 promotor, and provide a new way for astaxanthin production by plant genetic engineering.Methods: The Adketo gene w...Objective:To construct the plant expression vector including Adketo gene from Adonis aestivalis and E8 promotor, and provide a new way for astaxanthin production by plant genetic engineering.Methods: The Adketo gene was clonied from Adonis aestivalis by RT-PCR and the 1.1 kb tomato E8 gene was amplified by PCR. then cloned into T vector.After the preliminary identification by Microbial PCR, the recombinant T- vectors were subjected to sequence analysis. Cut the E8 promoter and Adketo gene and inserted into the plant binary expression vector pBI121to obtain the recombinant expression vector. And then to identify the vector by microbial PCR detection and enzyme digestion.Results: Sequencing results analysis showed the right E8 and Adketo gene sequences, the identification by microbial PCR and digestion with restriction enzymes proved that the recombinant vector had the inserts with expected length of target fragments.Conclusion: The plant expression vector containing Adketo gene driven by fruit specific promoter E8 was successfully constructed.展开更多
基金Hainan Natural Science Foundation Surface Project(20163077)Hainan Association of Science and Technology Young Science Talents Academic Innovation Project(HAST 201633).
文摘Objective:To construct the plant expression vector including Adketo gene from Adonis aestivalis and E8 promotor, and provide a new way for astaxanthin production by plant genetic engineering.Methods: The Adketo gene was clonied from Adonis aestivalis by RT-PCR and the 1.1 kb tomato E8 gene was amplified by PCR. then cloned into T vector.After the preliminary identification by Microbial PCR, the recombinant T- vectors were subjected to sequence analysis. Cut the E8 promoter and Adketo gene and inserted into the plant binary expression vector pBI121to obtain the recombinant expression vector. And then to identify the vector by microbial PCR detection and enzyme digestion.Results: Sequencing results analysis showed the right E8 and Adketo gene sequences, the identification by microbial PCR and digestion with restriction enzymes proved that the recombinant vector had the inserts with expected length of target fragments.Conclusion: The plant expression vector containing Adketo gene driven by fruit specific promoter E8 was successfully constructed.