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| The Constructing and Purification of Recombinant Human Fibroblast Growth Factor 8b Expressed Vector |
| HUANG Peng-huang1,2,3, WANG Ze1,2,3, TIAN Hai-shan2,4, ZHAO Hai-yang2, LI Hai-yan1,3, LI Xiao-kun1,2,4 |
1. ME Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agriculture University, Changchun 130021, China;
2. School of Pharmacy, Wenzhou Medical College, Wenzhou 325035, China;
3. School of Life Science, Jilin Agriculture University, Changchun 130021, China;
4. Zhejiang Gelusite Biology Technology Co. Ltd., Wenzhou 325000, China |
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Abstract FGF8b, which belongs to the fibroblast growth factor family, has been found to be associated with the regulation of growth and progression of hormonal cancers. Thus, it is believed that FGF8b could be a potential target in the treatment of hormonal cancers. To improve the production of recombinant hFGF8b to meet the increasing demand in basic research and clinical applications, an artificial gene encoding its mature peptide sequence was constructed and cloned into vector pET-3a. Then the recombinant proteins were expressed and presented in form of inclusion bodies in Escherichia coli BL21(DE3)pLysS. Expression, purification and renaturation conditions of the recombinant proteins were optimized. The preliminary biochemical characterization of FGF8b was further confirmed by Western blotting, and its mitogen activity was tested through MTT assay. In summary, this study has successfully acquired active recombinant FGF8b protein, which can be used for the further clinical development of embryology and the basic hormonal cancer researches.
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Received: 18 October 2012
Published: 25 January 2013
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