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Sadao Kato Tadao Kurata Shigeo Ishiguro Masao Fujimaki 《Bioscience, biotechnology, and biochemistry》2013,77(7):1759-1761
Bacillus circulans WL-12, a yeast and fungal cell wall lytic bacterium, secretes a variety of polysaccharide degrading enzymes into the culture medium. When β-1,3-glucanase was induced with pachyman, a β-1,3-glucose polymer obtained from the tree fungus Poria cocus Wolf, six distinct active molecules of the enzyme with different molecular weights were detected in the culture supernatant of this bacterium. Molecular cloning of one of the β,3-gIucanase genes into E. coli was achieved by transforming E. coli HB101 cells with recombinant plasmids composed of chromosomal DNA fragments prepared from B. circulans WL-12 and the plasmid vector pUC 19. A recombinant plasmid containing 4.4 kb of inserted DNA in the Pst I site of pUC 19, designated as pNT003, conferred the ability to degrade pachyman on E. coli cells. The presence of pNT003 was harmful for E. coli cells and caused cell lysis, especially at higher temperatures of cultivation. β,3-Glucanase activity detected in E. coli was mainly recovered in the periplasmic fraction when cell lysis did not occur. SDS-PAGE analysis revealed that the periplasmic fraction contained four active molecules of β-1,3-glucanase which corresponded to four of the six active molecules produced by B. circulans WL-12. 相似文献
74.
Tomohiko Mori Fumio Ibuki Setsuro Matsushita Tadao Hata 《Bioscience, biotechnology, and biochemistry》2013,77(9):1229-1235
The RNAs having template activities were extracted from soluble fraction of the cotyledons of soybean seeds. These were consisted of two major components, 9 s and 18 s (High molecular weight RNA, H-RNA). Both components have template activities in the E. coli S-30 system. H-RNA was found in the precipitate fraction when the so-called soluble fraction was centrifuged for 2 hr at 198,000×ɡ. H-RNA increased remarkably in kernels during ripening process and seems to be preserved in the seeds. 相似文献
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Aquporins are intrinsic membrane proteins that function as water channel to transport water and/or mineral nutrients across biological membranes. In this study, we aimed to clarify whether water structure can be changed by the presence of ceramics and whether such a change can be determined by aquaporin. First, we confirmed that ceramics could transform tap water into active tap water by increasing water permeability through aquaporin. We also found that this change in water permeability by treatment with ceramics occurred in distilled water. The distilled water was determined to exhibit the same aquaporin permeability as the original tap water. Our data indicate that the aquaporin permeability of water can be changed by severe physical shocks, such as slapping and sonication, which is consistent with the implication that the aquaporin permeability is closely related to the structure of the water. In this study, using aquaporins, we first reported that the treatment of water with ceramics can affect the structure of water, and the water can retain the structure for a given period under certain condition 相似文献
77.
Yokoi H Shimada A Carl M Takashima S Kobayashi D Narita T Jindo T Kimura T Kitagawa T Kage T Sawada A Naruse K Asakawa S Shimizu N Mitani H Shima A Tsutsumi M Hori H Wittbrodt J Saga Y Ishikawa Y Araki K Takeda H 《Developmental biology》2007,304(1):326-337
Medaka (Oryzias latipes) is a small freshwater teleost that provides an excellent developmental genetic model complementary to zebrafish. Our recent mutagenesis screening using medaka identified headfish (hdf) which is characterized by the absence of trunk and tail structures with nearly normal head including the midbrain-hindbrain boundary (MHB). Positional-candidate cloning revealed that the hdf mutation causes a functionally null form of Fgfr1. The fgfr1hdf is thus the first fgf receptor mutant in fish. Although FGF signaling has been implicated in mesoderm induction, mesoderm is induced normally in the fgfr1hdf mutant, but subsequently, mutant embryos fail to maintain the mesoderm, leading to defects in mesoderm derivatives, especially in trunk and tail. Furthermore, we found that morpholino knockdown of medaka fgf8 resulted in a phenotype identical to the fgfr1hdf mutant, suggesting that like its mouse counterpart, Fgf8 is a major ligand for Fgfr1 in medaka early embryogenesis. Intriguingly, Fgf8 and Fgfr1 in zebrafish are also suggested to form a major ligand-receptor pair, but their function is much diverged, as the zebrafish fgfr1 morphant and zebrafish fgf8 mutant acerebellar (ace) only fail to develop the MHB, but develop nearly unaffected trunk and tail. These results provide evidence that teleost fish have evolved divergent functions of Fgf8-Fgfr1 while maintaining the ligand-receptor relationships. Comparative analysis using different fish is thus invaluable for shedding light on evolutionary diversification of gene function. 相似文献
78.
An anchorage-dependent Wilms' tumor cell line, HFWT, has been found to be extremely susceptible to human natural killer (NK) cells. Here we established a transfectant of HFWT with the green fluorescence protein (GFP) gene, designated GHINK-1 cells, to apply to the activated NK cytotoxicity assay without radioisotope labeling. After being co-cultured with CD3 CD56+ NK cells, GHINK-1 cells released GFP into the medium. The intensity of the fluorescence from the released GFP correlated almost exactly with the radioactivity of a standard 51Cr-release assay performed with suspension-cultured K562 cells. Because it does not require separation of the remaining live target cells by centrifugation, the non-radioisotopic GFP release assay with GHINK-1 cells is a convenient alternative for monitoring human activated NK killing activity. 相似文献
79.
Uchida H Fujitani K Kawai Y Kitazawa H Horii A Shiiba K Saito K Saito T 《Bioscience, biotechnology, and biochemistry》2004,68(5):1004-1010
A new binding assay to investigate the mechanism of adhesion of lactic acid bacteria to the human intestine was established by the surface plasmon resonance technique using a biosensor BIACORE1000. Cells of 26 strains of the Lactobacillus acidophilus group as analytes were eluted onto a sensor chip on which were immobilized biotinylated A-trisaccharide polymer probes having human A-type antigen [(GalNAcalpha1-3(Fucalpha1-2)Gal)-] or human colonic mucin of blood type A (HCM-A) as ligands. In the first screening, high adhesive affinity to the A-trisaccharide BP-probe was observed in L. acidophilus OLL2769, L. crispatus JCM8778, LA205 and LA206. In the second screening, which used HCM-A, only L. acidophilus OLL2769 and L. crispatus JCM8778 were selected as adhesive strains with specific binding ability to human A-antigen. The results indicated that some strains of the L. acidophilus group could recognize and bind the sugar chain of A-antigen structure on HCM. 相似文献
80.
Montaño AM Kaitila I Sukegawa K Tomatsu S Kato Z Nakamura H Fukuda S Orii T Kondo N 《Human genetics》2003,113(2):162-169
Mucopolysaccharidosis IVA (MPS IVA) is caused by the deficiency of the lysosomal enzyme N-acetylgalactosamine-6-sulfate sulfatase encoded by the GALNS gene on chromosome 16. We describe, in detail, the clinical phenotype of five patients from three unrelated Finnish families and have characterized the disease-causing mutations in GALNS. Genotypes of the patients are D60N/A291T, D60N/W230X, and D60N/1374delT. Mutation 1374delT introduces premature termination of GALNS. Cells over-expressing the novel mutation W230X and A291T had no residual GALNS activity, whereas D60N gave 12.2% residual activity compared with the wild type. Co-transfection of D60N/A291T and D60N/W230X showed 5.5% and 6.7% of wild type activity, respectively. The precursor proteins of D60N and A291T were observed at 55 kDa and 57 kDa, respectively, whereas there was no detectable band in cells over-expressing W230X. At 55 degrees C, the mutant protein showed lower thermostability than the wild type protein at pH 3.8 and 7.0. The tertiary structural model of the GALNS protein revealed that aspartic acid at position 60 is located on the surface of the molecule, away from the active site. This makes it unlikely that the enzymatic function of the protein with D60N is severely impaired. On the other hand, A291 and W230 are localized near the active site. The molecular characteristics of the D60N mutation explain the attenuated clinical phenotype of the patients. 相似文献