Expression of a functional recombinant vascular endothelial growth factor 165 (VEGF₁₆₅) in Arabidopsis thaliana

Abstract

Objective Targeting the protein of interest to a particular tissue to achieve high-level expression is an important strategy to increase expression efficiency. The use of the plant seed oil body as a bioreactor can not only increase the amount of target protein, but also reduce the cost of downstream processing. Methods VEGF165 was expressed in Arabidopsis thaliana seeds via oilbody fusion technology. The pKO-VEGF165 vector was construted and transformed into A. thaliana seeds. T3 transgenic seeds was detected by SDS-PAGE and western blot methods. The cell activity was tested by MTT methods. Result The phaseolin promoter was used to drive seed-specific expression of the VEGF165 gene in transgenic A. thaliana. The coding region of VEGF165 was fused to the Arabidopsis oleosin sequence to target the protein to the oil bodies in the seeds of transgenic plants. The T-DNA region of recombinant plasmid pKO-VEGF165 was shifted to A. thaliana seeds via the floral-dip method. Protein was analyzed by electrophoresis and protein hybridization analyses. Finally, MTT assays showed that the oleosin-VEGF165 fusion protein played a part in the proliferation of HUVEC cells in vitro. Conclusion Oleosin-VEGF165 was successfully expressed and it had stimulated HUVEC cell proliferation activity.

Lipid Droplets from Plants and Microalgae: Characteristics, Extractions, and Applications

“…There are many kinds of growth factors, such as epidermal growth factor (EGF), fibroblast growth factor (FGF), insulin-like growth factor (IGF), interleukin (IL), nerve growth factor (NGF), etc. They can bind with the specific receptor of cell membranes and play an important role in regulating human immunity, hematopoiesis, nerve and hair follicle regeneration, wound healing, blood vessel formation, cell differentiation, and other aspects [ 111 , 116 , 117 , 118 , 119 , 120 , 121 ]. Plant LDs, as an expression system, provide a convenient, safe, and effective method for the production of these proteins [ 111 , 113 , 120 , 121 , 122 , 123 ].…”

“…They can bind with the specific receptor of cell membranes and play an important role in regulating human immunity, hematopoiesis, nerve and hair follicle regeneration, wound healing, blood vessel formation, cell differentiation, and other aspects [ 111 , 116 , 117 , 118 , 119 , 120 , 121 ]. Plant LDs, as an expression system, provide a convenient, safe, and effective method for the production of these proteins [ 111 , 113 , 120 , 121 , 122 , 123 ]. Human insulin-like growth factor 1 (hIGF-1) has been expressed in Arabidopsis thaliana seeds via oleosin fusion technology.…”