[Expression of biologically active recombinant arresten in Nicotiana tabacum].
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.
Li
Hongmin
H
School of Life Science, Northwest University, Xi'an 710069, China. hlhm2002@yahoo.com.cn
Hao
Jianguo
J
Jia
Jingfen
J
D016960:Agrobacterium tumefaciens / Q000235:genetics; D020533:Angiogenesis Inhibitors / Q000096:biosynthesis* / Q000235:genetics; D024141:Collagen Type IV / Q000096:biosynthesis* / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic
D020533:Angiogenesis Inhibitors; D024141:Collagen Type IV; D011994:Recombinant Proteins