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
We constructed an expression cassette of the organophosphorus pesticide degrading （opal） gene under the control of the E8 promoter. Then opd was transformed into tomato fruit using an agroinfiltration transient expr...We constructed an expression cassette of the organophosphorus pesticide degrading （opal） gene under the control of the E8 promoter. Then opd was transformed into tomato fruit using an agroinfiltration transient expression system. β-Glucuronidase （GUS） staining, reverse transcription-polymerase chain reaction （RT-PCR）, wavelength scanning, and fluorescent reaction were performed to examine the expression of the opd gene and the hydrolysis activity on coumaphos of organo- phosphorus hydrolase （OPH） in tomato fruit. The results show that the agroinfiltrated tomato fruit-expressed OPH had the maximum hydrolysis activity of about 11.59 Uhng total soluble protein. These results will allow us to focus on breeding transgenic plants that could not only enhance the degrading capability of fruit and but also hold no negative effects on pest control when spraying organophosphorus pesticides onto the seedlings in fields.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.
基金Project supported by the National Key Technology R&D Program of China(No.2007BAD59B06)the International Science and Technology Cooperation Program of China(No.2007DFA31260)
文摘We constructed an expression cassette of the organophosphorus pesticide degrading （opal） gene under the control of the E8 promoter. Then opd was transformed into tomato fruit using an agroinfiltration transient expression system. β-Glucuronidase （GUS） staining, reverse transcription-polymerase chain reaction （RT-PCR）, wavelength scanning, and fluorescent reaction were performed to examine the expression of the opd gene and the hydrolysis activity on coumaphos of organo- phosphorus hydrolase （OPH） in tomato fruit. The results show that the agroinfiltrated tomato fruit-expressed OPH had the maximum hydrolysis activity of about 11.59 Uhng total soluble protein. These results will allow us to focus on breeding transgenic plants that could not only enhance the degrading capability of fruit and but also hold no negative effects on pest control when spraying organophosphorus pesticides onto the seedlings in fields.
