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1.
A rapidly growing, maintainable, embryogenic suspension culture of Glycine max L. Merrill. has been generated. The culture consisted almost entirely of clumps of proliferating globular embryos with very little nonembryogenic tissues. The number and size of somatic embryo clumps were used to quantify growth of embryogenic tissues under various conditions. Initiation and proliferation of this embryogenic suspension culture were dependent on the inoculum, method of subculture, and composition of the subculture medium. Twenty to 50 mg of highly embryogenic, early-staged soybean tissue were inoculated into 35 ml of liquid culture medium containing 5 mg 1–1 2,4-D and either 15 mM glutamine or preferably 5 mM asparagine. Suspension cultures were subcultured at the same inoculum density every 4 weeks. The embryos matured and germinated following placement on solid media, resulting in consistent plant regeneration. 相似文献
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Yulia Makusheva Soo-Hyun Chung Aoi Akitsu Natsumi Maeda Takumi Maruhashi Xiao-Qi Ye Tomonori Kaifu Shinobu Saijo Haiyang Sun Wei Han Ce Tang Yoichiro Iwakura 《Experimental Animals》2022,71(3):288
Clec1A, a member of C-type lectin receptor family, has a carbohydrate recognition domain in its extracellular region, but no known signaling motif in the cytoplasmic domain. Clec1a is highly expressed in endothelial cells and weakly in dendritic cells. Although this molecule was reported to play an important role in the host defense against Aspergillus fumigatus by recognizing 1,8-dihydroxynaphthalene-melanin on the fungal surface, the roles of this molecule in un-infected animals remain to be elucidated. In this study, we found that Clec1a−/− mice develop milder symptoms upon induction of experimental autoimmune encephalomyelitis (EAE), an animal model for multiple sclerosis. The maximum disease score was significantly lower, and demyelination and inflammation of the spinal cord were much milder in Clec1a−/− mice compared to wild-type mice. No abnormality was detected in the immune cell composition in the draining lymph nodes and spleen on day 10 and 16 after EAE induction. Recall memory T cell proliferation after restimulation with myelin oligodendrocyte glycoprotein peptide (MOG35–55) in vitro was decreased in Clec1a−/− mice, and antigen presenting ability of Clec1a−/− dendritic cells was impaired. Interestingly, RNA-Seq and RT-qPCR analyses clearly showed that the expression of inflammatory cytokines including Il17a, Il6 and Il1b was greatly decreased in Clec1a−/− mice after induction of EAE, suggesting that this reduced cytokine production is responsible for the amelioration of EAE in Clec1a−/− mice. These observations suggest a novel function of Clec1A in the immune system. 相似文献
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Journal of Plant Research - The article Why is chlorophyll b only used in light-harvesting systems. 相似文献
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Akihito Tanaka Knut Woltjen Katsuya Miyake Akitsu Hotta Makoto Ikeya Takuya Yamamoto Tokiko Nishino Emi Shoji Atsuko Sehara-Fujisawa Yasuko Manabe Nobuharu Fujii Kazunori Hanaoka Takumi Era Satoshi Yamashita Ken-ichi Isobe En Kimura Hidetoshi Sakurai 《PloS one》2013,8(4)
The establishment of human induced pluripotent stem cells (hiPSCs) has enabled the production of in vitro, patient-specific cell models of human disease. In vitro recreation of disease pathology from patient-derived hiPSCs depends on efficient differentiation protocols producing relevant adult cell types. However, myogenic differentiation of hiPSCs has faced obstacles, namely, low efficiency and/or poor reproducibility. Here, we report the rapid, efficient, and reproducible differentiation of hiPSCs into mature myocytes. We demonstrated that inducible expression of myogenic differentiation1 (MYOD1) in immature hiPSCs for at least 5 days drives cells along the myogenic lineage, with efficiencies reaching 70–90%. Myogenic differentiation driven by MYOD1 occurred even in immature, almost completely undifferentiated hiPSCs, without mesodermal transition. Myocytes induced in this manner reach maturity within 2 weeks of differentiation as assessed by marker gene expression and functional properties, including in vitro and in vivo cell fusion and twitching in response to electrical stimulation. Miyoshi Myopathy (MM) is a congenital distal myopathy caused by defective muscle membrane repair due to mutations in DYSFERLIN. Using our induced differentiation technique, we successfully recreated the pathological condition of MM in vitro, demonstrating defective membrane repair in hiPSC-derived myotubes from an MM patient and phenotypic rescue by expression of full-length DYSFERLIN (DYSF). These findings not only facilitate the pathological investigation of MM, but could potentially be applied in modeling of other human muscular diseases by using patient-derived hiPSCs. 相似文献
5.
Ogawa A Murakami C Kamisuki S Kuriyama I Yoshida H Sugawara F Mizushina Y 《Bioorganic & medicinal chemistry letters》2004,14(13):3539-3543
A new isochroman derivative named pseudodeflectusin was isolated from a culture broth of Aspergillus pseudodeflectus. The structure was determined by spectroscopic means as 9-hydroxy-7-methyl-2-(methylethylidine)-furano[3,2-H]isochroman-3-one. This compound exhibited cytotoxicity for several human cancer cell lines from the stomach (NUGC-3), cervix (HeLa-S3), and peripheral blood (HL-60), but did not affect those from the lung (A549) or colon (DLD-1). The LD50 value of this compound for HL-60 cells was 39 microM. 相似文献
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Kuniko Akama Tomoe Horikoshi Atsushi Sugiyama Satoko Nakahata Aoi Akitsu Nobuyoshi Niwa Atsushi Intoh Yasutaka Kakui Michiko Sugaya Kazuo Takei Noriaki Imaizumi Takaya Sato Rena Matsumoto Hitoshi Iwahashi Shin-ichi Kashiwabara Tadashi Baba Megumi Nakamura Tosifusa Toda 《Biochimica et Biophysica Acta - Proteins and Proteomics》2010,1804(6):1272-1284
In mammalian spermiogenesis, sperm mature during epididymal transit to get fertility. The pig sharing many physiological similarities with humans is considered a promising animal model in medicine. We examined the expression profiles of proteins from boar epididymal caput, corpus, and cauda sperm by two-dimensional gel electrophoresis and peptide mass fingerprinting. Our results indicated that protein disulfide isomerase-P5 (PDI-P5) human homolog was down-regulated from the epididymal corpus to cauda sperm, in contrast to the constant expression of protein disulfide isomerase A3 (PDIA3) human homolog. To examine the functions of PDIA3 and PDI-P5, we cloned and sequenced cDNAs of pig PDIA3 and PDI-P5 protein precursors. Each recombinant pig mature PDIA3 and PDI-P5 expressed in Escherichia coli showed thiol-dependent disulfide reductase activities in insulin turbidity assay. Although PDIA3 showed chaperone activity to promote oxidative refolding of reduced denatured lysozyme, PDI-P5 exhibited anti-chaperone activity to inhibit oxidative refolding of lysozyme at an equimolar ratio. SDS-PAGE and Western blotting analysis suggested that disulfide cross-linked and non-productively folded lysozyme was responsible for the anti-chaperone activity of PDI-P5. These results provide a molecular basis and insights into the physiological roles of PDIA3 and PDI-P5 in sperm maturation and fertilization. 相似文献
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Yoshiki Morifuji Jian Xu Noriko Karasaki Kazuhiro Iiyama Daisuke Morokuma Masato Hino Akitsu Masuda Takumi Yano Hiroaki Mon Takahiro Kusakabe Jae Man Lee 《Molecular biotechnology》2018,60(12):924-934
Human α1-antitrypsin (AAT) is the most abundant serine proteinase inhibitor (serpin) in the human plasma. Commercially available AAT for the medications of deficiency of α1-antitrypsin is mainly purified from human plasma. There is a high demand for a stable and low-cost supply of recombinant AAT (rAAT). In this study, the baculovirus expression vector system using silkworm larvae as host was employed and a large amount of highly active AAT was recovered from the silkworm serum (~?15 mg/10 ml) with high purity. Both the enzymatic activity and stability of purified rAAT were comparable with those of commercial product. Our results provide an alternative method for mass production of the active rAAT in pharmaceutical use. 相似文献
9.
Summary The 5 and 3 flanking regions of the soybean glycinin gene, Gy1, responsible for expression in seeds, were analyzed by quantitative transient expression assay. The construct containing the -glucuronidase (uidA) reporter gene under the control of the 1.12 kb Gy1 promoter and 0.74 kb Gy1 terminator was introduced into immature soybean seeds and leaves by particle bombardment. To normalize the variability of introduction efficiency, a second reporter gene, firefly luciferase, was cobombarded as an internal standard, and relative activities (GUS/luciferase) were measured. There was a seed-specific -glucuronidase (GUS) expression, as observed by X-Gluc staining. Compared with the nopaline synthase gene (nos) terminator, the Gy1 terminator enhanced the level of expression in immature seeds, indicating that the terminator region of the glycinin gene is involved in the activation of the gene expression in these seeds. To identify cis-regulatory elements in the glycinin gene upstream sequence, deleted derivatives of the promoter were fused to the luciferase reporter gene. The expression could be measured with a higher accuracy, and constructs were introduced with the internal reporter uidA gene into immature seeds. The results suggest the presence of a positive regulatory element in the –620 to ––380 region of the Gy1 promoter. A deletion which eliminates the legumin box with its RY element led to increased relative activity, suggesting that this box is negatively regulating expression of the seed storage protein gene. Analysis of mutant promoters also suggest that the RY element involves negative regulation in seeds. This quantitative transient expression assay using particle bombardment provides a reliable system for the study of seed-specific gene expression in soybeans.Abbreviations GUS
-glucuronidase
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Gy1
glycinin AlaB2 gene
- CaMV
cauliflower mosaic virus
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nos
nopaline synthase gene
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uidA
-glucuronidase gene
- X-Gluc
5-bromo-4-chloro-3-indolyl glucuronide 相似文献
10.
Hideo Tsuji Akitsu Hitomi Ei-ichi Takahashi Motoi Murata Tatsuro Ikeuchi Kohtaro Yamamoto Satsuki Tsuji Tada-aki Hori 《Human genetics》1991,87(3):254-260
Summary Expression of distamycin A-inducible rare fragile sites by AT-specific DNA-ligands was examined in lymphoblastoid cell lines derived from heterozygous carriers for the fra(8)(q24), fra(16)(pl2), and fra(16)(q22) sites. The sensitivity of fragile site expression to the inducers was different at these fragile sites. The expression of fra(8)(q24) was induced markedly by Hoechst 33258, but not by distamycin A or berenil. An increased expression of fra(16)(p12) was found following treatment with Hoechst 33258 or berenil, but not with distamycin A. At fra(16)(q22), distamycin A markedly induced the fragile site, but Hoechst 33258 and berenil did not. Since their response to the different inducers was similar to that found in cultured lymphocytes, lymphoblastoid cell lines appear to retain their inherent properties. Although BrdUrd alone did nto induce any fragile sites, concomitant treatment with BrdUrd plus the inducer was synergistically effective in inducing all the fragile sites. An increased frequency of sister chromatid exchanges was observed at fra(16)(p12) following simultaneous treatment with BrdUrd and berenil, mainly when the site was expressed as an isochromatid gap. Thus, the induced fra (16)(pl2) site is a hot spot for the formation of sister chromatid exchanges, as found in other reported fragile sites. 相似文献