Production and secretion of human interleukin-18 in transgenic tobacco cell suspension culture |
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Authors: | Niti Sharma Tae-Geum Kim Moon-Sik Yang |
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Institution: | 1. Basic Science Research Institute, Chonbuk National University, 561-756, Jeonju, Korea 2. Division of Biological Sciences, Research Center for Bioactive Materials, Chonbuk National University, 561-756, Jeonju, Korea
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Abstract: | Interleukin-18 (IL-18), otherwise known as interferon-gamma-inducing factor (IGIF), is one of several well characterized and
important cytokines that contribute to host defenses. The complementary DNA (cDNA) of mature human interleukin-18 gene (hIL-18)
was fused with the signal peptide of the rice amylase 1A gene (Ramy1A) and introduced into the plant expression vector under
the control of a duplicated CaMV 355 promoter. The recombinant plasmid was transformed into tobacco (Nicotiana tabacum L. cv Havana) using theAgrobacterium-mediated transformation method. The integration of the hIL-18 gene into the genome of transgenic tobacco plants was confirmed
by polymerase chain reaction (PCR) amplification and its expression was observed in the suspension cells that were derived
from the transgenic plant callus by using Northern blot analysis. The hIL-18 protein was detected in the extracts of the transgenic
callus and in the medium of the transgenic tobacco suspension culture by using immunoblot analysis. Based upon enzyme-linked
immunosorbant assay (ELISA) results, the expression level of the hIL-18 protein approximated 166 μg/L in the suspension culture
medium. Bioassay results from the induction of interferon-γ from a KG-1 cell line indicated that the hIL-18 secreted into
the suspension culture medium was bioactive. |
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