Production system of recombinant protein hIL-32 in Pleurotus eryngii using Agrobacterium-mediated transformation
2009
Human recombinant IL-32 induces the production of large amounts of several proinflammatory cytokines and chemokines, even in macrophage cell lines by activation of NF-κB and MAPK. The ability of IL-32 to potentiate inflammation is not the sole action of this cytokine. IL- 32 stimulates prostaglandin (PG)E2 in human PBMCs, which are pivotal in inflammation. The development of recombinant protein consisting of IL-32 is of great medical and industrial importance. Agrobacterium strains GV3101 harboring pCAMBIA1304 include IL- 32 vectors were used to infect the mushroom fruiting body pieces. For transformation experiments, fruiting body gill tissue pieces were vacuum infiltrated with the suspension of induced bacteria until the air had been completely purged. Mushroom mycelium appeared at the margins of the tissue pieces after 9 to 14 days on selection medium with hygromycin at 50㎍/㎖. Transformants PCR analyses confirmed that the IL-32 gene was into the genome of Pleurotus eryngii. The present results demonstrating the usefulness of the transgenic technique s in the genetic manipulation and improvement of mushroom.
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