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Ibaraki, Japan

Baumann A.,Ohio State University | Barry J.,Ohio State University | Wang S.,Ohio State University | Wang S.,Chinese Academy of Agricultural Sciences | And 3 more authors.
Genetics | Year: 2010

Juvenile hormone (JH) is critical for multiple aspects of insect development and physiology. Although roles for the hormone have received considerable study, an understanding of the molecules necessary for JH action in insects has been frustratingly slow to evolve. Methoprene-tolerant (Met) in Drosophila melanogaster fulfills many of the requirements for a hormone receptor gene. A paralogous gene, germ-cell expressed (gce), possesses homology and is a candidate as a Met partner in JH action. Expression of gce was found to occur at multiple times and in multiple tissues during development, similar to that previously found for Met. To probe roles of this gene in JH action, we carried out in vivo gce over- and underexpression studies. We show by overexpression studies that gce can substitute in vivo for Met, alleviating preadult but not adult phenotypic characters. We also demonstrate that RNA interference-driven knockdown of gce expression in transgenic flies results in preadult lethality in the absence of MET. These results show that (1) unlike Met, gce is a vital gene and shows functional flexibility and (2) both gene products appear to promote JH action in preadult but not adult development. Copyright © 2010 by the Genetics Society of America. Source

Uchiyama K.,Utsunomiya University | Fujimoto H.,Japan National Institute of Infectious Diseases | Katsuma S.,University of Tokyo | Imanishi S.,Japan National Institute of Agrobiological Science | And 4 more authors.
In Vitro Cellular and Developmental Biology - Animal | Year: 2015

The Bombyx mori macula-like virus (BmMLV) is a member of the genus Maculavirus, family Tymoviridae, and contains a positive-sense single-stranded RNA genome. Previously, we reported that almost all B. mori-derived cell lines have already been contaminated with BmMLV via an unknown infection route. Since B. mori-derived cell lines are used for the baculovirus expression vector system, the invasion of BmMLV will cause a serious safety risk in the production of recombinant proteins. In this study, to determine the inactivation effectiveness of BmMLV, viruses were treated with various temperatures as well as gamma and ultraviolet (UV) light radiation. After these treatments, the virus solutions were inoculated into BmMLV-free BmVF cells. At 7 days postinoculation, the amount of virus in cells was evaluated by real-time reverse transcription PCR. Regarding heat treatment, conditions under 56°C for 3 h were tolerated, whereas infectivity disappeared after treatment at 75°C for 1 h. Regarding gamma radiation treatment, viruses were relatively stable at 1 kGy; however, their infectivity was entirely eliminated at a dose of 10 kGy. With 254 nm UV-C treatment, viruses were still active at less than 120 mJ/cm2; however, their infectivity was completely lost at greater than 140 mJ/cm2 UV-C radiation. These results provide quantitative evidence of the potential for BmMLV inactivation under a variety of physical conditions. © 2015 The Society for In Vitro Biology Source

Iwanaga M.,Utsunomiya University | Tsukui K.,Utsunomiya University | Uchiyama K.,Utsunomiya University | Katsuma S.,University of Tokyo | And 3 more authors.
Journal of Invertebrate Pathology | Year: 2014

We previously established the first Bombyx mori macula-like virus (BmMLV)-free cell line (BmVF cells) from a B. mori embryo. In this study, we evaluated the expression of recombinant proteins in BmVF cells using a B. mori nucleopolyhedrovirus (BmNPV)-derived expression vector. Our results showed that BmVF cells are susceptible to BmNPV, and both the promoter activity of the polyhedrin gene and the post-translated modifications of a recombinant protein are equivalent between BmMLV-negative BmVF and -positive BmN4 cells. These findings indicate that persistent infection with BmMLV has no discernible effect on BmNPV-mediated protein production in B. mori cells. © 2014 Elsevier Inc. Source

National Institute Of Agrobiological Science, Japan National Institute of Advanced Industrial Science, Technology and Preven Tec Inc. | Date: 2011-11-30

Providing a new method of treating inflammatory gastrointestinal disorders and a food product for the prevention or improvement of inflammatory gastrointestinal disorders. Prevention or treatment of inflammatory gastrointestinal disorders is possible by oral administration of recombinant IL-10 expressed in rice plant seeds.

Fujiwara Y.,Preventec Inc. | Aiki Y.,Preventec Inc. | Yang L.,Japan National Institute of Agrobiological Science | Takaiwa F.,Japan National Institute of Agrobiological Science | And 4 more authors.
Protein Expression and Purification | Year: 2010

Recombinant protein production system using transgenic rice grain offers many advantages in higher accumulation, preservation, lower production cost, ease of scale up and low risk of contamination by toxic materials. We developed a transgenic rice strain whose seeds accumulate human interleukin (IL)-10, a cytokine that suppresses inflammation-related immune responses. We also developed a method of extracting and purifying IL-10 from rice seeds. A biochemical crosslinking method was used to detect the biologically active noncovalent dimer of IL-10. This method was useful for developing efficient methods of refolding and purification. The purified IL-10 comprised only noncovalent dimers and showed higher activity than the commercial IL-10. The purified IL-10 had very low endotoxin contamination and is expected to have broad clinical application. © 2010 Elsevier Inc. All rights reserved. Source

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