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Molecular characterization of a cell wall-associated proteinase gene from Streptococcus lactis NCDO763 总被引:9,自引:0,他引:9
Streptococcus lactis NCDO763 harbours a plasmid designated pLP763. The cells harbouring pLP763 are able to grow to a higher density in milk because of their proteinase-positive phenotype (Prt+). The 6.2 kb HindIII-PstI fragment from pLP763 was found to be responsible for the Prt+ phenotype. The DNA fragment contains an incomplete large open reading frame (ORF). Further sequence analysis downstream from the PstI site revealed that the ORF consists of 5706 bases. It was found that the deduced amino acid sequence consisting of 1902 amino acid residues was extremely similar to that of the Wg2 proteinase, a serine protease from Streptococcus cremoris, suggesting that both genes were derived from a common ancestral gene. 相似文献
24.
Application of a novel apparatus, the quartz chemical analyzer, to the determination of endotoxin in blood. 总被引:2,自引:0,他引:2
A novel apparatus called a quartz chemical analyzer (QCA) has been developed using a quartz crystal resonator. This apparatus measures sample viscosity changes based on resonant frequency changes of the quartz crystal. The apparatus was used to determine bacterial endotoxin concentrations by monitoring the gelation reaction of Limulus amebocyte lysate. The QCA determined endotoxin concentrations with good accuracy and reproducibility in the range of 0.001-3 EU/ml for endotoxin standard (JP XII). For endotoxin determination in human whole blood and plasma samples, the inhibitory reaction was eliminated by pretreatment of a fourfold dilution at 60 degrees C and incubation for 30 min. There are many advantages of the QCA method compared with the turbidimetric and chromogenic methods. For example, QCA can measure sample viscosity changes with high sensitivity and accuracy because QCA detects minor resonant frequency changes and the frequency data give a numerical value for easy quantitation. QCA can examine turbid samples, and the required quantities of samples and reagents are small, since the quartz crystal detects sample viscosity changes directly. The endotoxin determination time may be shortened by raising the reaction temperature, and QCA can detect other types of coagulation reactions. 相似文献
25.
Cloning and characterization of motY, a gene coding for a component of the sodium-driven flagellar motor in Vibrio alginolyticus. 总被引:4,自引:3,他引:1 下载免费PDF全文
The bacterial flagellar motor is a molecular machine that couples proton or sodium influx to force generation for driving rotation of the helical flagellar filament. In this study, we cloned a gene (motY) encoding a component of the sodium-driven polar flagellar motor in Vibrio alginolyticus. Nucleotide sequence analysis revealed that the gene encodes a 293-amino-acid polypeptide with a single putative transmembrane segment that is very similar (94.5% identity) to the recently described MotY of V. parahaemolyticus. Their C-terminal domains were similar to the C-terminal domains of many peptidoglycan-interacting proteins, e.g., Escherichia coli MotB and OmpA, suggesting that MotY may interact with peptidoglycan for anchoring the motor. By using the lac promoter-repressor system, motY expression was controlled in V. alginolyticus cells. Swimming ability increased with increasing concentrations of the inducer isopropyl-beta-D-thiogalactopyranoside, and the swimming fraction increased after induction. These results are consistent with the notion that MotY is a component of the force-generating unit. V. alginolyticus motY complemented the motY mutation of V. parahaemolyticus. However, motY appeared to lack a region corresponding to the proposed motY promoter of V. parahaemolyticus. Instead, sequences similar to the sigma54 consensus were found in the upstream regions of both species. We propose that they are transcribed from the sigma54 -specific promoters. 相似文献
26.
Hideaki Matsuoka Yasushi Kazuno Takuji Horie Tomoo Homma Yasuyuki Nemoto 《Cytotechnology》1993,11(1):59-65
Possible roles of coexisting cells in inducing neurite growth from a nerve cell were studied. Nerve growth factor (NGF)-inducing neurite growth from PC12h-R (a cell line derived from cultured nerve cells) was investigated at various cell densities. At the cell density 102104 cells/ml neurites appeared even without NGF. In contrast, no neurite appeared without NGF in single cell culture. The neurite growth observed in plural cell culture without NGF was only partially inhibited by antibody to NGF receptor (Ab-NGFR). However, the effect of the used medium alone was mostly inhibited by Ab-NGFR. These results suggest that the neurite inducing potency of coexisting cells is via different sites than the NGF receptor.Abbreviations Ab-IgG-FITC
anti-mouse-IgG labeled with fluorescein isothiocyanate
- Ab-NF
monoclonal antibody to neurofilament 160 kD
- Ab-NGFR
monoclonal antibody to NGF receptor
- BDNF
brain-derived neurotrophic factor
- D-medium
medium for differentiation culture
- DMEM
Dulbecco's modified Eagle's medium
- M-medium
medium for multiplication culture
- NGF
nerve growth factor
- NGFR
NGF receptor
- NT-3
neurotrophin-3
- PC12
pheochromocytoma cell line
- PC12h-R
subclone of PC12
- Sup-D
supernatant of D-medium 相似文献
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H. Matsuoka H.-C. Yang T. Homma Y. Nemoto S. Yamada O. Sumita K. Takatori H. Kurata 《Applied microbiology and biotechnology》1995,43(1):102-108
Congo red was found to be feasible as a microscopic fluorescence indicator of hyphal growth at the single-hypha level. When 1 m Congo red was applied to mold of Aspergillus niger, the dye was found to a specific cell-wall component, chitin, without causing any inhibitory effect on hyphal growth. The bound Congo red emitted fluorescence at 614 nm. This binding reaction, however, proceeded more slowly than the growing speed of hypha. Consequently the fluorescence intensity was low at the apex where the surface area of the hypha was expanding rapidly. In contrast, as an apex where the growth was retarded, the fluorescence intensity became remarkably high. Therefore growing hyphae could be distinguished from non-growing hyphae by using Congo red. 相似文献
30.
S Kanegasaki Y Kojima M Matsuura J Y Homma A Yamamoto Y Kumazawa K Tanamoto T Yasuda T Tsumita M Imoto 《European journal of biochemistry》1984,143(2):237-242
Lipid A analogues were chemically synthesized based on the model structure recently revised, and biological activities of the analogues were tested. The analogue, (beta-1,6)-linked glucosamine disaccharide carrying ester-bound 3-hydroxytetradecanoic acids at 3 and 3' position of reducing and nonreducing glucosamine in addition to amide-bound 3-hydroxytetradecanoic acids and glycosidic-linked and ester-linked phosphate groups, showed much stronger activities for mediator inducing and immunomodulating as well as endotoxic activities than those exhibited by the previously synthesized analogues based on the old model. Among the activities tested, induction of interferon and tumor necrosis factor as well as mitogenicity, adjuvanticity and pyrogenicity were, however, not expressed so strongly as natural lipid A used as controls. In contrast, the analogue exhibited comparable activities to those of control lipid A in the test of lethal toxicity to mice and gelating activity of Limulus amebocyte lysate. Other synthetic analogues carrying a phosphate group showed comparable, slightly stronger or weaker activities depending on the test, but nonphosphorylated analogue exhibited no apparent or only very weak activities. 相似文献