Expression of biologically active recombinant arresten in Nicotiana tabacum.
- Author:
Hongmin LI
1
;
Jianguo HAO
;
Jingfen JIA
Author Information
1. School of Life Science, Northwest University, Xi'an 710069, China. hlhm2002@yahoo.com.cn
- Publication Type:Journal Article
- MeSH:
Agrobacterium tumefaciens;
genetics;
Angiogenesis Inhibitors;
biosynthesis;
genetics;
Collagen Type IV;
biosynthesis;
genetics;
Genetic Vectors;
genetics;
Plants, Genetically Modified;
genetics;
metabolism;
Recombinant Proteins;
biosynthesis;
genetics;
Tobacco;
genetics;
metabolism;
Transformation, Genetic
- From:
Chinese Journal of Biotechnology
2008;24(9):1520-1525
- CountryChina
- Language:Chinese
-
Abstract:
In this report, the biological activity of the recombinant Arresten expressed in Nicotiana tabacum was studied. The gene coding for the tumor angiogenesis inhibitor Arresten was PCR-amplified from the plasmid pCA and its plant expression vector named pCAMBIAarr was constructed by inserting the Arresten cDNA fragment into the NcoI/BstEII sites of the plant binary expression vector pCAMBIA1301. Then pCAMBIAarr was transferred into Agrobacterium tumefacien LBA4404 by the freeze-thaw method. The adventitious shoots and regenerated plants of Nicotiana tabacum with hygromycinB-resistance were obtained via Agrobacterium-mediated leaf disk transformation method. Southern hybridization, RT-PCR and Western blotting analysis showed that the Arresten cDNA was integrated into the genome of some of the regenerated plants and the recombinant Arresten was expressed with a molecular size of 26 kD. Recombinant Arresten purified from transgenic tobacco leaves had an anti-proliferative effect on bovine endothelial cells. We speculate that biologically active recombinant Arresten can be produced by using plants as bioreactors.
