Baculovirus-mediated Expression of a Gene for Trehalase of the Mealworm Beetle, Tenebrio molitor, in Insect Cells, SF-9, and Larvae of the Cabbage Armyworm, Mamestra brassicae |
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Authors: | Kenji Sato Miwa Komoto Toshitsugu Sato Hitoshi Enei Michihiro Michihiro Kobayashi Toshinobu Yaginuma |
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Institution: | a Iwate Biotechnology Research Center, 22-174-4 Narita, Kitakami, Iwate 024, Japan b School of Agricultural Sciences, Nagoya University, Chikusa, Nagoya 464-01, Japan |
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Abstract: | It is of interest to understand what kinds of physiological and biochemical changes occur in insects if the homeostasis of trehalose in the hemolymph is disrupted by the infection with a recombinant baculovirus containing a secretory-trehalase gene. For this purpose, two recombinant non-occluded Autographa california multicapsid nucleopolyhedroviruses (AcMNPVs), vTREVL and vERTVL, containing a trehalase cDNA of the mealworm beetle, Tenebrio molitor, were constructed. The trehalase cDNA was inserted in the sense orientation downstream of the polyhedrin promoter for vTREVL, and in the anitsense orientation for vERTVL. The active trehelase of T. molitor was found outside of cells when SF-9 cells or larvae of the cabbage armyworm, Mamestra brassicae, were infected with vTREVL. In the hemolymph of vTREVL-infected larvae, expression of the active trehelase was followed by disappearance of trehalose and appearance of glucose. However, the mortality time of virus-infected 5th instar larvae increased in the following order: AcMNPV C6 (wild-type virus) ≤ vERTVL < vTREVL. The symptoms (the browning and liquefying of the host body) of NPV infection were moderated considerably in vTREVL-infected larvae. |
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Keywords: | Trehelase Tenebrio molitor Baculovirus Autographa californica multicapsid nucleopolyhedrovirus Insect cell line Mamestra brassicae Trehalose Glucose Hemolymph |
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