Gene Synthesis, Protein Expression and Identification of VEGF189
CHEN Dan
WANG Hong
ZHU Zhongsong
XIANG Junjian
YANG Qin
ZHAO xin
DENG ning
Abstract：Objective To synthesis the gene for encoding VEGFI89 andexpress VEGFI89 protein in E. coli. Methods VEGF189 gene was synthesized using SOE PCR and cloned into pET-32a( + )vector. Then the recombinant expression plasmids were transfected into E. coli BL21 ( DE3 ) for expressing the fusion protein Trx-VEGF189. The expression products were identified by SDS-PAGE and Western blotting analysis respectively, and purified by HisTrapTM HP chromatography. Results DNA sequencing showed that VEGF189 gene was obtained successfully. The VEGF189 fusion protein was high level expressed in E. coli and the expressive prod-ucts were in the forms of inclusion bodies and soluble protein. The western blot analysis showed that the VEGF189 fusion protein could be recognized by anti-VEGF polyclonal antibody. The fusion proteins were purified effectively and the purity took up about 85%. Con-clusion The VEGF189 gene can be synthesized by SOE PCR and could be high level expressed in E. coil, which might pave the way for anti-tumor therapy by anti-VEGF antibody and tumor vaccine.
Keywords：VEGF189SOE PCRprokaryotic expression
Publication Date：2009-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 469-472 )