[Cloning, expression and identification of recombinant fusion protein GX1-rmhTNFalpha].

Abstract

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.

• Authors: Shan-shan Cao, Kai-chun Wu, Zhen Yan, Yi Wan, Yu Han, Li-na Zhao, Dai-ming Fan
• Journal: Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology (Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi) Vol. 22 Issue 3 Pg. 360-2 (May 2006) ISSN: 1007-8738 [Print] China
• PMID: 16643800 (Publication Type: Journal Article)

Chemical References

• Oligopeptides
• Peptides
• Peptides, Cyclic
• Recombinant Fusion Proteins
• Tumor Necrosis Factor-alpha
• cyclo(cysteinyl-glycyl-aspargyl-seryl-aspargyl-prolyl-lysyl-seryl-cysteine)

Topics

• Blotting, Western
• Cloning, Molecular
• Drug Delivery Systems
• Escherichia coli (genetics)
• Gene Expression
• Genetic Vectors
• Humans
• Molecular Sequence Data
• Oligopeptides (chemistry, genetics, metabolism)
• Peptides (genetics, metabolism)
• Peptides, Cyclic
• Recombinant Fusion Proteins (genetics, metabolism, therapeutic use)
• Temperature
• Tumor Necrosis Factor-alpha (genetics, metabolism)