The Role of Bacterial Vectors in the Expression of Human IFN-م Gene
2003
Moeen Rezakhanlou, A., Immunology and Hematology Dept., School of Medicine, Hamedan University of MedicalSciences, Hamedan, Iran | Mahboudi, F., Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran | Maghsoudi, N., Biotechnology Research Center, Malek Ashtar Industrial University, Tehran, Iran | Habibi, G.R., Razi Vaccine and Serum Research Institute, Tehran, Iran
The Interferon gamma (IFN-م) 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 E.coli. The transcription of IFN-م mRNA was determined by northern blot analysis. The level of expression of human IFN-م 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-م under the control of promoters of pET32a, pQE30 and pSKA-IBA plasmids was detected by SDS-GAGE and western blotting.
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