Biotechnology and Bioprocess Engineering:BBE

  • 제11권2호
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  • Pages.154-159
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  • 2006
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  • 1226-8372(pISSN)
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  • 1976-3816(eISSN)
한국생물공학회 (The Korean Society for Biotechnology and Bioengineering)
권호 표지

Production and Secretion of Human Interleukin-18 in Transgenic Tobacco Cell Suspension Culture

  • Sharma, Niti (Basic Science Research Institute, Chonbuk National University) ;
  • Kim, Tae-Geum (Division of Biological Sciences, Research Center for Bioactive Materials, Chonbuk National University) ;
  • Yang, Moon-Sik (Division of Biological Sciences, Research Center for Bioactive Materials, Chonbuk National University)
  • 발행 : 2006.04.30
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초록

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 35S promoter. The recombinant plasmid was transformed into tobacco (Nicotiana tabacum L. cv Havana) using the Agrobacterium-mediated transformation method. The integration of the hlL-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 hlL-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 hlL-18 protein approximated $166{\mu}g/L$ in the suspension culture medium. Bioassay results from the induction of $interferon-{\gamma}$ from a KG-1 cell line indicated that the hlL-18 secreted into the suspension culture medium was bioactive.

키워드

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