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Molecular cloning and expression of novel fibroblast growth factor-2 conjugated with immunodominant domains of pseudomonas exotoxin

Haghighatfard, H. and Samaei, N.M. and Farazmandfar, T. and Hakemi, M.G. and Yamchi, A. and Jadidi-Niaragh, F. and Yazdani, Y. (2015) Molecular cloning and expression of novel fibroblast growth factor-2 conjugated with immunodominant domains of pseudomonas exotoxin. Biomedical and Pharmacology Journal, 8 (2). pp. 1195-1200.

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Abstract

Angiogenesis is very important in cancer growth and metastasis. Basic fibroblast growth factor (bFGF) as one of the most important angiogenesis factors is an attractive target for cancer vaccine. Due to low immunogenicity, it cannot stimulate an effective immune response. Theoretically, pseudomonas exotoxin (PE) as a potent immunogenic carrier protein when fused to low immunogenic antigens such as bFGF significantly increased immunogenicity of it. In this study, we tried to molecular cloning and expression of bFGF conjugated with immunodominant domains of pseudomonas exotoxin. The coding sequence of fusion protein composed of bFGF linked to PE domains 1b and 2 using EAAAK poly linker. The KDEL sequence was also used in C-terminal coding sequence. It was synthesized and expressed using recombinant DNA technology in the bacterial expression system. Expression of recombinant protein verified using SDS-PAGE and western blot analyses. Finally, it purified using Ni-affinity chromatography. The band close to 37 kDa in SDS-PAGE and western blot analyses was aligned completely to designed sequence. Purified recombinant protein also showed as a clear single band near to 37 kDa.

Item Type: Article
Additional Information: cited By 0
Uncontrolled Keywords: fibroblast growth factor 2; Pseudomonas exotoxin, affinity chromatography; alpha helix; angiogenesis; Article; cancer cell; circulation; gene expression; human; immune response; immunogenicity; large scale production; molecular cloning; nonhuman; polyacrylamide gel electrophoresis; recombinant DNA technology; Western blotting
Subjects: مقالات نمایه شده محققین دانشگاه در سایت ,Web of Science ,Scopus
Divisions: معاونت تحقیقات و فناوری
Depositing User: GOUMS
Date Deposited: 10 Sep 2016 07:24
Last Modified: 09 Jan 2017 09:34
URI: http://eprints.goums.ac.ir/id/eprint/4777

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