Paper Information

Journal:   ARCHIVES OF RAZI INSTITUTE   June 2003 , Volume - , Number 55; Page(s) 71 To 80.
 
Paper: 

THE ROLE OF BACTERIAL VECTORS IN THE EXPRESSION OF HUMAN IFN- Γ GENE

 
 
Author(s):  MOEEN REZAKHANLOU A., MAHBOUDI F.*, MAGHSOUDI N., HABIBI GHOLAM REZA
 
* Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
 
Abstract: 

The Interferon gamma (IFN-γ) gene 1 was isolated from phytohemagglutinin stimulated peripheral blood mononuclear cells of a healthy individual blood donor using RT-PCR technique. The gene was cloned under the control of the different promoters expression vectors such as pET32a (Novagen), pQE30 (QIAGEN), pSKA-IBA (Strep-Tag), pRSET (Invitrogen), and then expressed in Ecoli. The transcription of IFN- γ mRNA was determined by northern blot analysis. The level of expression of human IFN-γ in pRSET vector under the control of T7 promoter was determined by laser-based densitometry of SDSPAGE and found more than 26% of total bacterial protein. The expression was confirmed by western blotting. The expression of IFN-γ under the control of promoters of pET32a, pQE30 and pSKA-IBA plasmids was detected by SDS-GAGE and western blotting.

 
Keyword(s): INTERFERON-Γ, EXPRESSION, E. COLI, NORTHERN BLOTTING
 
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