CLONING OF HUMAN IFN-λ1 (IL-29) FROM MONOCYTE DERIVED DCS AND ITS EXPRESSION IN HEK 293 T

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AMIR KALVANGH P.; EBTEKAR M.; AZADMANESH K.; HARTOONIAN C.; MAHDAVI M.

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Journal: ARAK MEDICAL UNIVERSITY JOURNAL (AMUJ) Year:2011 | Volume:14 | Issue:4 (57) Page(s): 69-78

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Title

CLONING OF HUMAN IFN-λ1 (IL-29) FROM MONOCYTE DERIVED DCS AND ITS EXPRESSION IN HEK 293 T

Author(s)

Keywords

INTERFERON LAMBDA 1Q2

CLONINGQ2

EXPRESSIONQ2

PCRQ2

Abstract

Background: Type III Interferon (IFN) is a novel member of the interferon family, which contains three ligands: IFN-l1 (IL-29), IFN-l2 (IL-28A) and IFN-l3 (IL-28B).These three ligands use the same unique heterodimeric receptor composed of CRF2-12 (IFN-l-R1/IL-28Ra) and CRF2-4 (IL10- R-b) chains which are completely different from type I & type II IFN receptors. IFNsl exhibit several features such as antiviral activity, antiproliferative activity, immunomodulatory activity and in vivo antitumour activity. In this work we aimed to clone the ogene of IFN-l1 obtained from dendritic cells and assess protein production in eukaryotic EXPRESSION vector.Materials and methods: in thid experimental study, total RNA was extracted from monocyte derived dendritic cells stimulated with 100 ng/ml of LPS. cDNA was synthesized from total RNA.Then cDNA of IFN-l1 was amplified by PCR with specific primers and cloned into the PTZ57R/Tvector in the E.coli (DH5a). This was subsequently subcloned into plasmid pcDNA3.1+, using KpnI and BamHI restriction endonucleases. After tranfection into HEK293 T, EXPRESSION of protein was tested by sandwich-ELISA method.Results: The DNA sequence of the insert was identical to the published sequences encoding IFN-l1 in GeneBank. It was demonstrated that IFN-l1 gene was markedly transcribed in transfected cells. EXPRESSION of IFN-l1 in HEK293 T cells was confirmed by sandwich ELISA.Conclusion: Successful CLONING and EXPRESSION IFN-l1 can be the first step for more production and further investigation about other activities of this cytokine and provides grounds for research on obtaining new therapeutic approaches for cancer, viral, autoimmune and allergic disease and designing more effective vaccines.

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APA: Copy

AMIR KALVANGH, P., EBTEKAR, M., AZADMANESH, K., HARTOONIAN, C., & MAHDAVI, M.. (2011). CLONING OF HUMAN IFN-λ1 (IL-29) FROM MONOCYTE DERIVED DCS AND ITS EXPRESSION IN HEK 293 T. ARAK MEDICAL UNIVERSITY JOURNAL (AMUJ), 14(4 (57)), 69-78. SID. https://sid.ir/paper/69450/en

Vancouver: Copy

AMIR KALVANGH P., EBTEKAR M., AZADMANESH K., HARTOONIAN C., MAHDAVI M.. CLONING OF HUMAN IFN-λ1 (IL-29) FROM MONOCYTE DERIVED DCS AND ITS EXPRESSION IN HEK 293 T. ARAK MEDICAL UNIVERSITY JOURNAL (AMUJ)[Internet]. 2011;14(4 (57)):69-78. Available from: https://sid.ir/paper/69450/en

IEEE: Copy

P. AMIR KALVANGH, M. EBTEKAR, K. AZADMANESH, C. HARTOONIAN, and M. MAHDAVI, “CLONING OF HUMAN IFN-λ1 (IL-29) FROM MONOCYTE DERIVED DCS AND ITS EXPRESSION IN HEK 293 T,” ARAK MEDICAL UNIVERSITY JOURNAL (AMUJ), vol. 14, no. 4 (57), pp. 69–78, 2011, [Online]. Available: https://sid.ir/paper/69450/en

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