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Published on: October 30, 2012
[Cloning, expression and identification of recombinant fusion protein GX1-rmhTNFalpha]
Shan-shan Cao1, Kai-chun Wu, Zhen Yan
1State Key Laboratory of Cancer Biology, Institute of Digestive Diseases, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. caoshansahn80@yahoo.com.cn
Aim:
To construct prokaryotic vector GX1-rmhTNF containing neovascular-targeting peptide GX1 and human TNFalpha and produce the GX1-rmhTNF protein.
Methods:
We have previously obtained a specific phage peptide CGNSNPKSC (GX1) binding to vasculature of human gastric cancer. The GX1-rmhTNFalpha vector was constructed by merging sequence of neovascular-targeting peptide GX1 with N-terminal of new recombined human TNFalpha (rmhTNFalpha) using gene engineering methods. The expression of the fusion protein GX1-rmhTNFalpha in E. coli was induced by temperature. The expression of GX1-rmhTNFalpha was detected by SDS-PAGE and Western blot.
Results:
A novel protein with expected molecular mass about 18,000 was found after SDS-PAGE and gel staining. The expressed product showed a good binding ability to anti-TNFalpha monoclonal antibody.
Conclusion:
The prokaryotic vector GX1-rmhTNF was constructed and the expression of GX1-rmhTNF protein was successfully induced, which was helpful for further purification of GX1-rmhTNF protein.
