The Constructing And Expression Of Recombinant Human Fibroblast Growth Fator-8a High Level Expressed Vector

The member of the Fibroblast Growth Factor familiy,Fibroblast Growth Factor-8, isoform a(FGF8a)is accepted as a crucial organizing signal during the development of vertebrate embryos. Furthermore, the FGF8a is also efficient in healing organic wound, culturing the neural stem cells and induction of dopaminergic neurons. So it is suggested that the potential application is huge for FGF8a to benefit the cliniacal treatment . Our study is a part of the rhFGF8a project, which is cooperated by Guangzhou Biotechnology Co.,Ltd and New Zealand Crown Research Institute. The former studies had constructed a rhFGF8a-vector with low expression level and demonstrated the efficacy of rhFGF8a. So we recombined the rhFGF8a again and constructed two high level expressed vectors. Using the techniques of molecular cloning and DNA point mutation skill, the mature recombinant human FGF8a cDNA was inserted into the pET22b(+) and pET28a(+), which was transformed into Eschericha coli Rosetta(DE3) and BL21 (DE3)pLysS. The induction experiment and protein analyse have revealed that the expression levels of the two new vectors are much higher than the original rhFGF8a vector. The cell culture in vitro experiment has proved the activity of rhFGF8a protein. In summary, our study have constructed two new vector to express the rhFGF8a protein in high level. They could be available for large-scale industrial production and play an important role in the biological and clinical research of rhFGF8a.