Biopolym. Cell. 2003; 19(2):179-184.
Molecular and Cell Biotechnologies
Biosynthesis of human basic fibroblast growth factor in soluble form in Escherichia coli and its purification
- PSRS "Biotechnolog"
150, Akademika Zabolotnogo Str., Kyiv, Ukraine, 03680
Basic fibroblast growth factor (βFGF, also known as FGF-2) is an important modulator of cell differentiation, proliferation, migration, migration, angiogenesis and other activities in a variety of cell types. The recombinant human βFGF is considered as a potential medicine for the treatment of vascular and some other diseases. In this work the production of human bFGF of 146 a. a. length, in E. coli cells, using the phage lambda, and its purification have been elaborated. The method of recombinant protein production includes the phage lambda infection of the BL21 (DE3) cells, containing a plasmid with a target gene under the T7 phage promoter. The process results in lysis of the producer strain cells and releasing newly synthesized βFGF into the cell medium, where it accumulates in a soluble form (approximately 60 mg/l). The target protein has been purified to homogeneity not less than 98 % by a one-step procedure using heparin-Sefarose affinity chromatography. According to the biological tests βFGF obtained promotes angiogenesis in chick embryo chorioallantoic membrane.
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