Cloning and Expression of Human Gamma-Interferon cDNA in E. coli

Document Type: Research Paper

Authors

1 National Research Center for Genetic Engineering and Biotechnology (NRCGEB), P. O. Box: 14155-6343, Tehran, Iran.

2 Biotechnology Research Center, P.O. Box: 19395-1949, Tehran, Iran.

Abstract

Prior to the production of human gamma interferon using recombinant DNA technology, it had been produced
mainly upon mitogenic induction of lymphocytes in very low amounts, which evidently hampered
its characterization and its medical applications. The recombinant gamma interferons produced in larger
quantities in prokaryotic systems retain their biological activities, and can be used clinically in the treatment
of various viral, neoplastic and immunosuppressed conditions or diseases. In this study, a cDNA
sequence coding for human gamma interferon was synthesized from mRNA template extracted from
induced human T lymphocytes. The cDNA was then amplified by PCR, cloned in an expression vector,
and transformed into Escherichia coli. The polypeptide produced through the expression of this DNA
sequence in E. coli showed immunological and chemical properties resembling authentic human IFN-γ.

Keywords