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Paper Information

Journal:   BIOLOGICAL SCIENCES (DANISH-I ZISTI-I IRAN)   SPRING 2008 , Volume 3 , Number 1; Page(s) 43 To 56.
 
Paper: 

EVALUATING THE EFFECT OF RARE CODON REPLACEMENT WITH OPTIMAL CODONS ON EXPRESSION LEVEL OF HUMAN BASIC FIBRIBLAST GROWTH FACTOR (HBFGF) IN ESCHERICHIA COLI

 
 
Author(s):  ABEDI KHALILABAD M.*, MIRZAHOSSEINI H., JAVADINIA GH.
 
* DEPARTMENT OF BIOLOGY, SCIENCE & RESEARCH BRANCH, ISLAMIC AZAD UNIVERSITY, TEHRAN, IRAN
 
Abstract: 

The Expression level of recombinant proteins in Escherichia coli can be affected by several factors. It has been observed that codon replacement at Translation Initiation Region (TIR), critically changes the heterologous expression in E. coli. This study aims to evaluate the effect of replacing rare codons with optimal codons in TIR on the expression level of human basic Fibriblast Growth Factor (hbFGF) in E.coli. For this purpose a long primer with 5 altered codons for the relative amino acid were designed to construct the TIR in a different way. The altered gene was then cloned in an expression vector and new construct was transformed into 3 different strains of E.coli. The induction of protein expression in these strains exhibited a significant change in expression levels which reemphasized the role of TIR codons in the efficiency of protein expression in E.coli. In conclusion the results suggest that nucleotide sequence in TIR has a key role in the level of hbFGF expression in E. coli.

 
Keyword(s): HUMAN BASIC FIBRIBLAST GROWTH FACTOR (HBFGF), CODON REPLACEMENT ,TRANSLATION INITIATION REGION (TIR), RARE CODON, OPTIMAL CODON
 
References: 
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