This paper should be cited as: Haghighatfard, H. Yazdani, Y.
Designing and Construction of Recombinant Plasmid Consisting of Basic
Fibroblast Growth Factor and Immunodominant
Fragments of Pseudomonas Exotoxin
Haghighatfard, H. (BSc)1, Yazdani,
Y. (PhD)2
1. MSc Student of Medical Biotechnology,
Faculty of Advanced Medical Technologies, Golestan
University of Medical Science, Gorgan, Iran
2. Assistant Professor of Immunology, Infectious Diseases Research
Center and Laboratory Science Research Center, Golestan
University of Medical Sciences, Gorgan, Iran
Abstract
Background
and Objective: the inhibition of tumor-associated
angiogenesis can significantly reduce the
tumor proliferation. The basic fibroblast growth factor
(bFGF), an important angiogenic
factor, is considered as a potential therapeutic target for cancer therapy. The
purpose of this study was evaluating, designing and construction of new
recombinant
Material
and Methods: Different types of peptide linker, codon
adaptation index (CAI) and adding signal
peptide were considered in designing of immunogenic coding sequence. After
software evaluation, the recombinant DNA molecule was ordered in the puc57
cloning vector. Then, coding sequence inserted into the multiple cloning site
of pET28-a plasmid. Finally, PCR and enzymatic digestion
tests were done for evaluation of recombinant expression vector.
Results: Optimization of DNA sequence, codon adaptation index (CAI) increased from 0.69 to 0.83 and GC content decreased from 61 to 54.77. The presence of 1214-bp PCR product
and 1029-bp one obtaining from enzymatic digestion confirmed the correction of
the cloning process.
Conclusion: According to the previous studies, it is the first
work for designing, optimizing and synthesis of recombinant DNA consisting of bFGF and immunodominant epitopes
of Pseudomonas toxin.
Keywords:
Tumor angiogenesis, immunodominant epitopres of
Pseudomonas toxin, Fibroblast growth factor 2, DNA 2 software
Corresponding Author: Yazdani, Y.
Email: yaghobyazdani59@yahoo.com
Received: 7 Jun 2014
Revised: 12 Sep 2014
Accepted: 15 Sep 2014
This paper should be cited as: Haghighatfard
H, Yazdani Y. [Designing and Construction of
Recombinant Plasmid Consisting of Basic Fibroblast Growth Factor and Immunodominant Fragments of Pseudomonas Exotoxin]. mljgoums. 2015; 9(1): 45-52 [Article
in Persian]