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排序方式: 共有177条查询结果,搜索用时 15 毫秒
21.
Takeo Sato Shugo Maekawa Shigetaka Yasuda Yutaka Sonoda Etsuko Katoh Takanari Ichikawa Miki Nakazawa Motoaki Seki Kazuo Shinozaki Minami Matsui Derek B. Goto Akira Ikeda Junji Yamaguchi 《The Plant journal : for cell and molecular biology》2009,60(5):852-864
Plants are able to sense and respond to changes in the balance between carbon (C) and nitrogen (N) metabolite availability, known as the C/N response. During the transition to photoautotrophic growth following germination, growth of seedlings is arrested if a high external C/N ratio is detected. To clarify the mechanisms for C/N sensing and signaling during this transition period, we screened a large collection of FOX transgenic plants, overexpressing full‐length cDNAs, for individuals able to continue post‐germinative growth under severe C/N stress. One line, cni1‐D (carbon/nitrogen insensitive 1‐dominant), was shown to have a suppressed sensitivity to C/N conditions at both the physiological and molecular level. The CNI1 cDNA encoded a predicted RING‐type ubiquitin ligase previously annotated as ATL31. Overexpression of ATL31 was confirmed to be responsible for the cni1‐D phenotype, and a knock‐out of this gene resulted in hypersensitivity to C/N conditions during post‐germinative growth. The ATL31 protein was confirmed to contain ubiquitin ligase activity using an in vitro assay system. Moreover, removal of this ubiquitin ligase activity from the overexpressed protein resulted in the loss of the mutant phenotype. Taken together, these data demonstrated that CNI1/ATL31 activity is required for the plant C/N response during seedling growth transition. 相似文献
22.
Kondou Y Higuchi M Takahashi S Sakurai T Ichikawa T Kuroda H Yoshizumi T Tsumoto Y Horii Y Kawashima M Hasegawa Y Kuriyama T Matsui K Kusano M Albinsky D Takahashi H Nakamura Y Suzuki M Sakakibara H Kojima M Akiyama K Kurotani A Seki M Fujita M Enju A Yokotani N Saitou T Ashidate K Fujimoto N Ishikawa Y Mori Y Nanba R Takata K Uno K Sugano S Natsuki J Dubouzet JG Maeda S Ohtake M Mori M Oda K Takatsuji H Hirochika H Matsui M 《The Plant journal : for cell and molecular biology》2009,57(5):883-894
Ectopic gene expression, or the gain-of-function approach, has the advantage that once the function of a gene is known the gene can be transferred to many different plants by transformation. We previously reported a method, called FOX hunting, that involves ectopic expression of Arabidopsis full-length cDNAs in Arabidopsis to systematically generate gain-of-function mutants. This technology is most beneficial for generating a heterologous gene resource for analysis of useful plant gene functions. As an initial model we generated more than 23 000 independent Arabidopsis transgenic lines that expressed rice fl-cDNAs (Rice FOX Arabidopsis lines). The short generation time and rapid and efficient transformation frequency of Arabidopsis enabled the functions of the rice genes to be analyzed rapidly. We screened rice FOX Arabidopsis lines for alterations in morphology, photosynthesis, element accumulation, pigment accumulation, hormone profiles, secondary metabolites, pathogen resistance, salt tolerance, UV signaling, high light tolerance, and heat stress tolerance. Some of the mutant phenotypes displayed by rice FOX Arabidopsis lines resulted from the expression of rice genes that had no homologs in Arabidopsis . This result demonstrated that rice fl-cDNAs could be used to introduce new gene functions in Arabidopsis. Furthermore, these findings showed that rice gene function could be analyzed by employing Arabidopsis as a heterologous host. This technology provides a framework for the analysis of plant gene function in a heterologous host and of plant improvement by using heterologous gene resources. 相似文献
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Taishi Hashiguchi Takanari Kobayashi Duriya Fongmoon Ajaya Kumar Shetty Shuji Mizumoto Nobuyuki Miyamoto Toshikazu Nakamura Shuhei Yamada Kazuyuki Sugahara 《Biochimica et Biophysica Acta (BBA)/General Subjects》2011
Background
Chondroitin sulfate (CS) is a ubiquitous component of the cell surface and extracellular matrix and its sugar backbone consists of repeating disaccharide units: D-glucuronic acid (GlcUA)β1-3N-acetyl-D-galactosamine (GalNAc). Although CS participates in diverse biological processes such as growth factor signaling and the nervous system's development, the mechanism underlying the functions is not well understood.Methods
CS was isolated from ray fish cartilage, an industrial waste, and its structure and neurite outgrowth-promoting (NOP) activity were analyzed to investigate a potential application to nerve regeneration.Results
The major disaccharide unit in the CS preparation was GlcUA-GalNAc(6-O-sulfate) (61.9%). Minor proportions of GlcUA-GalNAc(4-O-sulfate) (27.0%), GlcUA(2-O-sulfate)-GalNAc(6-O-sulfate) (8.5%), and GlcUA-GalNAc (2.7%) were also detected. The preparation showed NOP activity in vitro, and this activity was suppressed by antibodies against hepatocyte growth factor (HGF) and its receptor c-Met, suggesting the involvement of the HGF signaling pathway in the expression of the in vitro NOP activity of the CS preparation. The specific binding of HGF to the CS preparation was also demonstrated by surface plasmon resonance spectroscopy.Conclusions and general significance
The NOP activity of CS from ray cartilage was demonstrated to be expressed through the HGF signaling pathway, suggesting that ray cartilage CS may be useful for studying the cooperative function of CS and HGF. 相似文献25.
Optimized protocols and plasmids for in vivo cloning in yeast 总被引:1,自引:0,他引:1
Kitazono AA 《Gene》2011,484(1-2):86-89
Saccharomyces cerevisiae has proven a valuable system for the construction of plasmids via gap repair or in vivo cloning. The method allows cloning with superior accuracy and without the need to use restriction enzymes. However, despite its remarkable efficiency, the process may occasionally require the screening of large number of candidates. We have previously reported that by simply using shuttle plasmids that allow blue/white selection in Escherichia coli, it is possible to pre-select for positive clones. Here, we demonstrate that the same strategy can be used to assemble plasmids from several ectopic DNA fragments, which are all introduced in yeast cells by a simple transformation step. Further, to facilitate the subcloning of the fragment cloned into other targeting or expression vectors, the multi-cloning sites of three shuttle plasmids have been extended to include fifteen new restriction enzyme recognition sites. 相似文献
26.
Brain pericytes are an important constituent of neurovascular unit. They encircle endothelial cells and contribute to the
maturation and stabilization of the capillaries in the brain. Recent studies have revealed that brain pericytes play pivotal
roles in a variety of brain functions, such as regulation of capillary flow, angiogenesis, blood brain barrier, immune responses,
and hemostasis. In addition, brain pericytes are pluripotent and can differentiate into different lineages similar to mesenchymal
stem cells. The brain pericytes are revisited as a key player to maintain brain function and repair brain damage. 相似文献
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29.
Ai Sasou Takanari Shigemitsu Yuhi Saito Manami Tanaka Shigeto Morita Takehiro Masumura 《Plant cell reports》2016,35(6):1287-1295
Key message
Prolamin–GFP fusion proteins, expressed under the control of native prolamin promoters, were localized in specific layers of PB-Is. Prolamin–GFP fusion proteins were gradually digested from outside by pepsin digestion.Abstract
In rice seed endosperm, protein body type I (PB-I) has a layered structure consisting of prolamin species and is the resistant to digestive juices in the intestinal tract. We propose the utilization of PB-Is as an oral vaccine carrier to induce mucosal immune response effectively. If vaccine antigens are localized in a specific layer within PB-Is, they could be protected from gastric juice and be delivered intact to the small intestine. We observed the localization of GFP fluorescence in transgenic rice endosperm expressing prolamin–GFP fusion proteins with native prolamin promoters, and we confirmed that the foreign proteins were located in specific layers of PB-Is artificially. Each prolamin–GFP fusion protein was localized in specific layers of PB-Is, such as the outer-most layer, middle layer, and core region. Furthermore, to investigate the resistance of prolamin–GFP fusion proteins against pepsin digestion, we performed in vitro pepsin treatment. Prolamin–GFP fusion proteins were gradually digested from the peripheral region and the contours of PB-Is were made rough by in vitro pepsin treatment. These findings suggested that prolamin–GFP fusion proteins accumulating specific layers of PB-Is were gradually digested and exposed from the outside by pepsin digestion.30.