Shingo Nagaya
Nara Institute of Science and Technology
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Featured researches published by Shingo Nagaya.
Plant and Cell Physiology | 2010
Shingo Nagaya; Kazue Kawamura; Atsuhiko Shinmyo; Ko Kato
To express a foreign gene in plants effectively, a good expression system is required. Here we describe the identification of a transcriptional terminator that supports increased levels of expression. The terminators of several Arabidopsis genes were examined in transfected Arabidopsis T87 protoplasts. The heat shock protein 18.2 (HSP) terminator was the most effective in supporting increased levels of expression. The HSP terminator increases mRNA levels of both transiently and stably expressed transgenes approximately 2-fold more than the NOS (nopaline synthase) terminator. When combined with the HSP terminator, a translational enhancer increased gene expression levels approximately 60- to 100-fold in transgenic plants.
Molecular Genetics and Genomics | 2001
Shingo Nagaya; Kazuya Yoshida; Ko Kato; K. Akasaka; Astuhiko Shinmyo
Abstract. Specialized DNA sequences known as insulators protect genes from both the positive and negative influences of nearby chromatin. Many insulators have been identified in various species; however, few function in multiple species. We have shown that an insulator from the Ars (arylsulfatase) gene of the sea urchin Hemicentrotus pulcherrimus functions in plant cells. Normally, expression of an introduced chimeric GUS gene is inactivated in approximately 30% of transformed tobacco BY2 clones. Transgenes containing the Ars insulator, however, were expressed in all transformed tobacco BY2 cells. The insulator did not affect the copy number, the chromosomal position of transgene integration or maximum expression levels. These results suggest that the insulator functions to suppress the variation normally associated with transgene expression in tobacco BY2 cells.
Journal of Bioscience and Bioengineering | 2000
Shingo Nagaya; Yuhta Nakai; Ko Kato; Masami Sekine; Kazuya Yoshida; Atsuhiko Shinmyo
Strong promoters are required under several culture conditions for effective transgene expression in tobacco BY2 cells. We have isolated the promoter fragments of 4 genes exhibiting high homology to those of Arabidopsis thaliana 108C1T7 (unknown function) and F1-ATPase-delta, alcohol dehydrogenase and pectin esterase genes from a genomic DNA library of BY2 cells. Two of the four genes were strongly expressed during every phase of growth of BY2 cells, and the other two were expressed only during the stationary phase. Each of the promoter fragments was ligated to the GUS reporter gene and introduced into the chromosome of BY2 cells by Agrobacterium-mediated transformation. Growth-phase-dependent expression of the GUS gene was reproduced under the control of all 4 promoters observed with the original genes. Significantly higher expression was observed under the control of Nt108p during every phase of cell growth and under the control of NtADHp and NtPESp during the stationary phase than that under the control of the CaMV35S promoter.
Plant Biotechnology | 2011
Yoshimi Oshima; Nobutaka Mitsuda; Masaru Nakata; Tsuyoshi Nakagawa; Shingo Nagaya; Ko Kato; Masaru Ohme-Takagi
Plant and Cell Physiology | 2005
Shingo Nagaya; Ko Kato; Yuka Ninomiya; Rie Horie; Masami Sekine; Kazuya Yoshida; Atsuhiko Shinmyo
Plant and Cell Physiology | 2007
Yuichi Kodama; Shingo Nagaya; Atsuhiko Shinmyo; Ko Kato
Jarq-japan Agricultural Research Quarterly | 2005
Ryutaro Aida; Shingo Nagaya; Kazuya Yoshida; Sanae Kishimoto; Michio Shibata; Akemi Ohmiya
Biotechnology and Bioengineering | 1998
Atsuhiko Shinmyo; Takeshi Shoji; E. Bando; Shingo Nagaya; Y. Nakai; Ko Kato; Masami Sekine; Kazuya Yoshida
Plant Biotechnology | 2007
Yuichi Kodama; Shingo Nagaya; Atsuhiko Shinmyo; Ko Kato
Archive | 2000
Atsuhiko Shinmyo; Kazuya Yoshida; Ko Kato; Shingo Nagaya; Michio Shibata; Ryutaro Aida