科研之友
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摘要
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: VEGF(165 )was expressed in Arabidopsis thaliana seeds via oilbody fusion technology. The pKO-VEGF(165) 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 VEGF(165) gene in transgenic A. thaliana. The coding region of VEGF(165) 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-VEGF(165) 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-VEGF(165) fusion protein played a part in the proliferation of HUVEC cells in vitro. @@@ Conclusion: Oleosin-VEGF(165) was successfully expressed and it had stimulated HUVEC cell proliferation activity.
- 出版日期2019-6
- 单位吉林农业大学
