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Production of functional human vascular endothelial growth factor165 in transgenic rice cell suspension cultures

Paper ID Volume ID Publish Year Pages File Format Full-Text
17169 42648 2014 6 PDF Available
Title
Production of functional human vascular endothelial growth factor165 in transgenic rice cell suspension cultures
Abstract

•We constructed transgenic rice cell suspension culture producing human VEGF165.•The expression of rhVEGF165 was determined by Northern and Western blot analyses.•The accumulated rhVEGF165 protein in the culture medium was 19 mg/L.•The rhVEGF165 was purified using a heparin HP column.•Its biological activity was tested on human umbilical endothelial cells (HUVECs).

Vascular endothelial growth factors (VEGFs) are secreted by tumor cells and other cells exposed to hypoxia, and play a critical role in the development and differentiation of the vascular system. In this study, we investigated the production of functional recombinant human VEGF165 (rhVEGF165) in transgenic rice cell suspension culture. Complementary DNA was synthesized from human leukemia HL60 cells and cloned into expression vectors under the control of the rice α-amylase 3D (RAmy3D) promoter. The rice seed (Oryza sativa L. cv. Dongjin) was transformed with this recombinant vector by the Agrobacterium mediated method and the integration of the target gene into the plant genome was confirmed by genomic PCR. The expression of rhVEGF165 in the rice cells was determined by Northern blot and Western blot analyses. The accumulated rhVEGF165 protein in the culture medium was 19 mg/L after 18 days of culturing in a sugar-free medium. The rhVEGF165 was purified using a heparin HP column and its biological activity was tested on human umbilical vein endothelial cells (HUVECs). The purified rhVEGF165 significantly increased the proliferative activity of the HUVECs. Therefore, it was demonstrated that functional rhVEGF165 could be produced using transgenic rice suspension culture vector under the control of the RAmy3D promoter.

Keywords
Rice cell suspension culture; Recombinant human VEGF165; RAmy3D promoter; Heparin HP column
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Production of functional human vascular endothelial growth factor165 in transgenic rice cell suspension cultures
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 63, September 2014, Pages 58–63
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us