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Heterogeneous Expression and Purification of the Wheat VRN1 K-Box Domain Suggest the Formation of a Tetramer of the VRN1 Protein

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American Journal of Plant Sciences 2021年第7期12卷 1002-1010页

作者： takafumi itoh Ayane Kamiya Moe Kimura Koji Murai Faculty of Bioscience and Biotechnology Fukui Prefectural University Fukui Japan Department of Sustainable Agri-Culture Fukui Prefectural University Fukui Japan

In cereal species such as wheat (Ttiticum aestivum) and barley (Hordeum vulgare), many studies have indicated that VERNALIZATION 1 (VRN1) functions as a flowering promoter, which activates florigen gene expression. The wheat florigen gene Wheat FLOWERING LOCUS T (WFT, which is identical to VRN3) is an integrator of the vernalization, photoperiod and autonomous pathways in wheat flowering, and the WFT expression is correlated with the VRN1 expression. VRN1 encodes an APETALA1/FRUITFULL-like MADS-box transcription factor which expression is induced by vernalization, leading to flowering thorough up-regulation of

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Efficient Production of L-Theanine Using Immobilized Recombinant <i>Escherichia coli</i>Cells Expressing a Modified γ-Glutamyltranspeptidase Gene from <i>Pseudomonas nitroreducens</i>

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Advances in Biological Chemistry 2020年第6期10卷 157-171页

作者： Putthapong Phumsombat Chiharu Sano Hiroki Ikezoe Junji Hayashi takafumi itoh Takao Hibi Mamoru Wakayama Department of Biotechnology College of Life Sciences Ritsumeikan University Shiga Japan Graduate School of Technology Industrial and Social Sciences Tokushima University Tokushima Japan Department of Bioscience and Biotechnology Fukui Prefectural University Fukui Japan

L-Theanine (γ-glutamylethylamide) is a naturally occurring amino acid derivative known to have several beneficial physiological effects as a diet supplement, and to give an umami taste when used as a food additive. The compound is industrially produced by γ-glutamyltranspeptidase from Pseudomonas nitroreducens (PnGGT). Using recombinant PnGGT, we have shown previously that Trp385, Phe417, and Trp525 are key amino acid residues for recognition of acceptor substrates at the PnGGT active site. Here, we demonstrate that a recombinant W525D mutant of PnGGT produces L-theanine from ethylamine and L-glutamine more efficiently than wild-type PnGGT, attributable to an increased ratio of transfer activity to hydrolysis activity. An efficient production of L-theanine was achieved by immobilizing Escherichia coli cells expressing the W525D PnGGT mutant (E. coli-W525D) using 2% alginate as the supporting material. The highest L-theanine production using immobilized E. coli-W525D, representing a conversion rate of 90%, was achieved in optimal reaction conditions of pH 10, 40°C, and a substrate molar ratio of L-glutamine to ethylamine of 1:10. The immobilized E. co

关键词： L-Theanine γ-Glutamyltranspeptidase Pseudomonas nitroreducens Escherichia coli Immobilization

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Leukemogenic Fusion Gene (p190 BCR-ABL) Transduction into Hematopoietic Stem/Progenitor Cells in the Common Marmoset

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Open Journal of Blood Diseases 2012年第1期2卷 1-10页

作者： Yan Dong Seiichiro Kobayashi Yamin Tian Manabu Ozawa takafumi Hiramoto Kiyoko Izawa Yuansong Bai Yasushi Soda Erika Sasaki Toshio itoh Yoshiro Maru Satoshi Takahashi Kaoru Uchimaru Naoki Oyaizu Arinobu Tojo Chieko Kai Kenzaburo Tani Central Institute for Experimental Animals Kawasaki Japan Department of Laboratory Medicine Research Hospital Institute of Medical Science the University of Tokyo Tokyo Japan Department of Molecular Genetics Medical Institute of Bioregulation Kyushu University Fukuoka Japan Department of Pharmacology Tokyo Woman’s Medical University Japan Division of Molecular Therapy Institute of Medical Science the University of Tokyo Tokyo Japan Laboratory Animal Research Center Institute of Medical Science the University of Tokyo Tokyo Japan.

Patients with Philadelphia chromosome (p190 BCR-ABL fusion gene)-positive acute lymphoblastic leukemia have a poor prognosis despite intensive therapeutic *** this study, we attempted to develop a leukemia nonhuman primate model that mimics various human systems. Hematopoietic stem/progenitor cells in the common marmoset were transduced with a lentiviral vector containing the p190 BCR-ABL fusion gene by ex vivo transduction or in vivo direct bone marrow injection. In the latter model, BCR-ABL gene expression was maintained for more than one and a half years. One marmoset unexpectedly developed myelofibrosis-like disease. However, none of the marmosets have developed leukemia to date. In conclusion, we successfully achieved sustained p190 BCR-ABL gene expression in vivo. However, a genetic mutation in addition to p190 BCR-ABL may be required for the malignant transformation of hematopoietic stem/progenitor cells in the common marmoset during the short observation period. This novel in vivo approach will help develop a marmoset leukemia model in the future.

关键词： Leukemia Lentiviral Vector Myelofibrosis Common Marmoset

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