Cloning And Expression Of Human Basic Fibroblast Growth Factor (hbFGF) CDNA

Basic fibroblast growth factor (bFGF) is a member of the fibroblast growth factor family and also a mitogenic, angiogenic and neurotrophic factor. Human basic fibroblast growth factor (hbFGF) has been implicated in a multitude of physiologic and pathologic processes, including proliferation, differentiation, angiogenesis, wound healing and tumor growth. Recently, more and more hospitals and beauty parlors began to cure some symptoms with hbFGF as the investigation was advanced. But the problem raised is the high price for hbFGF so that it can't be widespread. At the same time, hbFGF is not stable with high bioactivities. Therefore it is very urgent to create new industrial technique to produce stable hbFGF with low cost and high quantity.For the moment, E coli expression system normally is used to produce recombinant hbFGF (rhbFGF) protein. But hbFGF has low solubility and stability and is not stable in solutions, the dimer or multimer of hbFGF is easily formed. hbFGF also has low output in the prokaryotic expression system[1]. On the one hand, there is four Cys amino acids at 34, 78, 96 and 101 sites, which easily form dimer by disulfide bond between different molecules to reduce hbFGF's bio-activities. On the other hand, there is some codons in hbFGF gene which are rare in E.coli. That causes the low expression efficiency and quantity.In order to find the method to improve hbFGF expression and investigate the correlation between its structure and function, the cDNA encoding hbFGF needs to be cloned and expressed in E coli. Then it will be...