Summary
An anti-metastatic polypeptide, bifunctional-domain (Cell I -Hep I) recombinant polypeptide of human fibronectin, was expressedin E. coli and purified. The expression level was found to be about 20%– 30 % of the total cell proteins. In BL21 (DE3)/T7, anE. coli expressing system, lactose can be used as an inducer to substitute IPTG thereby reducing the cost by several hundredfold and it is suitable for the large-scale preparation of recombinant FN polypeptide. Cell I -Hep 1 fragment is an alkaline polypeptide. In BL21(DE3)/T7 expressing system, better isolation was achieved if DEAE-52, instead of CM-52, was used for ion-exchange chromatography. The purified product was obtained after heparin-agarose affinity chromatography following ion-exchange chromatography.
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References
Kornblihtt A, Umezawa K, Vibe-Pedersen Ket al. Primary structure of human fibronectin: differential splicing may generate at least 10 polypeptides from a single gene. The EMBO J, 1985, 4(7): 1755
Kimizuka F, Taguchi Y, Ohaate Yet al. Production and Characterization of functional domains of human fibronection expressed in Escherichia coli. J Biochem, 1991, 110: 284
Saiki I, Makabe T, Yoneda Jet al. Inhibitory effect of fibronection and its recombinant polypeptides on the adhesion of metastatic melanoma cells to laminin. Jpn J Cancer Res, 1991, 82: 1112
Matsumoto Y, Saiki Y, Makabe Tet al. Inhibitory effect of antimetastatic fusion polypeptide of human fibronectin on tumor cell adhesion to. extracellular matrices. Jpn J Cancer Res, 1991, 82: 1130
Yoneda J, Saiki I, Kobayashi Het al. Inhibitory effect of recombinant fibronectin polypeptides on the adhesion of liver-metastatic lymphoma cells to hepaticsinusoidal endothe-lial cells and tumorinvasion. Jpn J Cancer Res, 1994, 85: 723
Saiki I, Murata J, Makabe Tet al. Inhibition of lung metastasis by synthetic and recombinant fragments of human fibronectin with functional domains. Jpn J Cancer Res, 1990, 81: 1003
Saiki I, Matsumoto Y, Murata Jet al. Recombinant fusion polypeptide with Celland Heparin-binding domains of fibronectin inhibits liver metastasis of L5178Y-ML25 lymphoma cells. Jpn J Cancer Res, 1991, 82: 1120
Feng Zuohua, Zhang Guimei, Li Donget al. Construction of expressing plasmids of recombinant FN polypeptides with bifunctional-domain and the characterization of the products expressed inE. coli. J Tongji Med Univ, 1996, 16(2): 70
Studier F W, Moffatt B A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression cloned geges. J Mol Biol, 1986, 189: 113
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This project was supported by grants from National Natural Science Foundation of China and Science Foundation of the Ministry of Public Health (No. 39370783).
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Guimei, Z., Zuohua, F., Dong, L. et al. Investigation on the purification and expression of cell I — Hep I recombinant FN polypeptide inE. Coli . Journal of Tongji Medical University 16, 134–138 (1996). https://doi.org/10.1007/BF02908792
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DOI: https://doi.org/10.1007/BF02908792