02865nam a2200289Ia 4500 MX-MdCICY 20260521091716.0 CICY B-19922 A hydroponic cultivation system for rapid high-yield transient protein expressioin in Nicotiana plants under laboratory conditions Journal of Southern Medical University, 32(6), p.772-777, 2012 Objective: To develop a hydroponic Nicotiana cultivation system for rapid and high-yield transient expression of recombinant proteins under laboratory conditions. Methods: To establish the hydroponic cultivation system, several parameters were examined to define the optimal conditions for the expression of recombinant proteins in plants. We used the green fluorescent protein (GFP)and the geminiviral plant transient expression vector as the model protein/expression vector. We examined the impact of Nicotiana species, the density and time of Agrobacterium infiltration, and the post-infiltration growth period on the accumulation of GFP. The expression levels of GFP in Nicotiana leaves were then examined by Western blotting and ELISA. Results: Our data indicated that a hydroponic Nicotiana cultivation system with a light intensity of 9000 LX/layer, a light cycle of 16 h day/8 h night, a temperature regime of 28 degrees celsius; day/21 degrees celsius; night, and a relative humidity of 80 percent could support the optimal plant growth and protein expression. After agroinfiltration with pBYGFPDsRed.R/LBA4404, high levels of GFP expression were observed in both N. benthamiana and N. tobaccum (cv. Yuyan No.5)plants cultured with this hydroponic cultivation system. An optimal GFP expression was achieved in both Nicotiana species leaves 4 days after infiltration by Agrobacterium with an OD(600)of 0.8. At a given time point, the average biomass of N. tobaccum (cv. Yuyan No.5)was significantly higher than that of N. benthamiana. The leaves from 6-week-old N. benthamiana plants and 5-week-old N. tobaccum (cv. Yuyan No.5)plants could be the optimal material for agroinfiltration. Conclusion: We have established a hydroponic cultivation system that allows robust growth of N. benthamiana and N. tobaccum (cv. Yuyan No.5)plants and the optimal GFP expression in the artificial climate box. NICOTIANA PLANTS TRANSIENT EXPRESSION HYDROPONIC CULTIVATION GREEN FLUORESCENT PROTEIN Mo, Q. Mai, R. Yang, Z. Chen, M. Yang, T. Lai, H. Zhou, X. https://drive.google.com/file/d/1xMB9pSMJwnd5QyVuiNqH-2mDmWsp8cM8/view?usp=drivesdk Para ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx Loc REF1 250602s9999 xx |||||s2 |||| ||und|d 29967 29967 0 0 Loc 0 0 F1 CICY CICY RE 2025-06-25 0 B-19922 2025-06-25 16:24:39 2025-06-25 REF1