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81.
Cloning, nucleotide sequence and expression of a hemolysin gene of Clostridium septicum 总被引:4,自引:0,他引:4
Abstract A genomic library of Clostridium septicum NCTC547 strain was made in Escherichia coli by means of λgt10. The DNA insert of a hemolysin-positive (Hly+ ) λ-clone was transferred into pUC19. The resulting plasmid, pCS21, confers a Hly+ phenotype on E. coli . Crude lysates of E. coli (pCS21) possessed a strong lytic activity on human erythrocytes and also a lethal effect on mice, characteristic of an α toxin. Nucleotide sequence analysis revealed that the insert DNA (5.2 kb) in pCS21 included at least one open reading frame of 1380 bp. The coding frame for hemolysin was predicted to be 1329 bp in size and to encode a protein of 49.8 kDa. It coincided with the molecular mass (48 kDa) of the α toxin secreted by C. septicum . Taken together, the data indicated that plasmid pCS21 indeed encoded an α toxin gene of C. septicum . 相似文献
82.
Stable isotopic structure of aquatic ecosystems 总被引:1,自引:0,他引:1
Isotopic, biogeochemical and ecological structure can provide a new dimension for understanding material flows, and the simultaneous
function and structure of an ecosystem. Distributions ofδ
13C andδ
15N for biogenic substances in the Nanakita river estuary involving Gamo lagoon in Japan were investigated to construct isotope
biogeochemical and ecological structure for assessing fate and transfer of organic matter, and food web structure. The isotopic
framework of the ecosystem was successfully described in aδ
15N–δ
13C map. In this estuary the variations of isotope ratios of biogenic substances were clearly explained by the mixing of land-derived
organic matter, and marine-derived organic matter.
A trophic-level effect of15N enrichment was clearly observed. Organisms were classified into three groups depending upon the contribution of land-derived
organic matter in a food chain. Almost all biota except mollusca in the lagoon depend on organic matter of marine origin.
The contributions of both land and marine organic matter were comparable for mollusca in the lagoon. 相似文献
83.
84.
Noriyuki Suka Yoshinobu Shinohara Yasushi Saitoh Hisato Saitoh Kohei Ohtomo Masahiko Harata Edward Shpigelman Shigeki Mizuno 《Genetica》1993,88(2-3):93-105
About 65% of DNA in the chicken W chromosome has been shown to consist ofXhoI andEcoRI family repetitive sequences. These sequences showed remarkable delay in the electrophoretic mobility at low temperature
on a polyacrylamide gel. Three dimensional structures of the 0.7-kbXhoI and the 1.2-kbEcoRI family repeating units were estimated to be irregular solenoids using a computer program based on wedge angles of all the
16 dinucleotide steps. Fluorescencein situ hybridization demonstrated that these two family sequences were localized in a major heterochromatic body in an interphase
nucleus. Incorporation of bromodeoxyuridine into the W chromosome in the synchronous culture of MSB-1 cells occurred about
1 h later than the peak of S phase. The chromatin structure formed alongXhoI andEcoRI family sequences was suggested to be different from the total chromatin or chromatin containing the β-actin gene sequence
in that the linker DNA lengths of the former were significantly longer. Fractionation of theHaeIII-digested MSB-1 nuclei yielded a chromatin fraction in whichXhoI family sequences were partially enriched. Several DNA-binding proteins showing higher affinity for theXhoI family sequence were present in this fraction. 相似文献
85.
Yasushi Sato Munetaka Sugiyama Ryszard J. Górecki Hiroo Fukuda Atsushi Komamine 《Planta》1993,189(4):584-589
In a culture system in which single cells isolated from the mesophyll of Zinnia elegans L. differentiate to tracheary elements (TEs), two inhibitors of phenylalanine ammonia-lyase (EC 4.3.1.5), L-α-aminooxy-β-phenylpropionic acid (AOPP) at 10 μM inhibited lignification without reducing the number of TEs formed. These inhibitors caused intracellular changes in peroxidase (EC 1.11.1.7) activities. The inhibitors increased the activity of peroxidases bound to the cell walls and especially the activity of peroxidase bound ionically to the cell walls. In contrast, the activity of extracellular peroxidase decreased. There were five isoenzymes, P1-P5, in the ionically bound peroxidase of cultured Zinnia cells. Among the isoenzymes, P4 and P5 appeared to be specific for TE differentation. Treatment with AOPP and AIP resulted in increases in the activities of P2, P4 and P5 isoenzymes, with the most prominent increase in P5 activity. The addition of lignin precursors, including coniferyl alcohol, to the AOPP-treated cells restored lignification, and suppressed the alteration of peroxidase isoenzyme patterns caused by AOPP. The relationship between the wall-bound peroxidases and lignification during TE differentiation is discussed in the light of these results. 相似文献
86.
Inactivation of erythropoietin receptor function by point mutations in a region having homology with other cytokine receptors. 总被引:21,自引:11,他引:10
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The cytoplasmic domain of the erythropoietin receptor (EpoR) contains a region, proximal to the transmembrane domain, that is essential for function and has homology with other members of the cytokine receptor family. To explore the functional significance of this region and to identify critical residues, we introduced several amino acid substitutions and examined their effects on erythropoietin-induced mitogenesis, tyrosine phosphorylation, and expression of immediate-early (c-fos, c-myc, and egr-1) and early (ornithine decarboxylase and T-cell receptor gamma) genes in interleukin-3-dependent cell lines. Amino acid substitution of W-282, which is strictly conserved at the middle portion of the homology region, completely abolished all the functions of the EpoR. Point mutation at L-306 or E-307, both of which are in a conserved LEVL motif, drastically impaired the function of the receptor in all assays. Other point mutations, introduced into less conserved amino acid residues, did not significantly impair the function of the receptor. These results demonstrate that conserved amino acid residues in this domain of the EpoR are required for mitogenesis, stimulation of tyrosine phosphorylation, and induction of immediate-early and early genes. 相似文献
87.
Kazuyoshi Masuda Shunji Nagata Koichiro Hirano Yasushi Takagishi Hidematsu Hirai 《Microbiology and immunology》1993,37(2):165-167
This paper describes an attempt to effectively induce antibody-dependent cell-mediated cytotoxicity (ADCC) in nude mice. A monoclonal antibody against α-fetoprotein, 80G, coadministered with spleen cells from other nude mice bearing HuH-7N (xenograft of human hepatoma cell line, HuH-7) significantly suppressed the growth of HuH-7N as compared to treatment with 80G alone. 80G with spleen cells from normal nude mice also had some suppressive effect. In contrast, no effect was observed with each spleen cells alone as well as 80G alone. These results suggest that further supply of effector cells could enhance ADCC activity in nude mice. 相似文献
88.
Spatial structures of landforms as constants affecting vegetation patterns are discussed based on analyses of hilly land regions, mostly in the vicinity of Sendai, northeast Japan. The lower part of a hillslope is characterized by relatively active processes of soil erosion, landslides and slope failure. It supports a plant community different from that on the upper part of the hillslope. These two parts are termed the lower hillslope and the upper hillslope, respectively. The upper hillslope consists of valley heads with no stream water nor stream channels. Since a valley head is comprised of several micro-scale landform units, an upper hillslope can be subdivided into these landform units. Plant communities vary in their species compositions and structures as well as in some other ecological characteristics paralleling changes in their respective micro-scale landform units. However, the variations in species composition within upper hillslopes are not as extreme as those between the upper and lower hillslopes. xx]Papers presented at the Vth INTECOL Congress at YOKO-hama 1990. 相似文献
89.
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
90.
Kazuyoshi Masuda Shunji Nagata Koichiro Hirano Yasushi Takagishi Hidematsu Hirai 《生物化学与生物物理学报:疾病的分子基础》1993,1182(2):128-132
This study was carried out to clarify the reason for elevation of serum α-fetoprotein (AFP) level of nude mice bearing hepatoma cells after treatment with monoclonal antibodies (MoAbs) to AFP. MoAbs to AFP showed no effect on the cumulative amounts of AFP secreted from human hepatoma cell line, HuH-7, in vitro. However, the treatment of nude mice bearing HuH-7N cells (HuH-7 xenograft) with MoAbs to AFP led to elevation of the serum AFP level in spite of the fact that the growth curve of HuH-7N cells was similar to that for PBS treatment. This apparent elevation of the serum AFP level is thought to be due to the slow elimination of AFP-MoAb immune complexes with little lattice structure from circulation, but not the enhancement of AFP secretion of HuH-7N cells. Thus, when using a MoAb alone or MoAb-drug conjugate, the serum AFP level should only be cautiously used as a tumor marker for evaluating the targeting immunotherapy. 相似文献