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61.
62.
Cloning and characterization of two catA genes in Acinetobacter lwoffii K24. 总被引:3,自引:0,他引:3
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S I Kim S H Leem J S Choi Y H Chung S Kim Y M Park Y K Park Y N Lee K S Ha 《Journal of bacteriology》1997,179(16):5226-5231
Two novel type I catechol 1,2-dioxygenases inducible on aniline media were isolated from Acinetobacter lwoffii K24. Although the two purified enzymes, CD I1 and CD I2, had similar intradiol cleavage activities, they showed different substrate specificities for catechol analogs, physicochemical properties, and amino acid sequences. Two catA genes, catA1 and catA2, encoding by CD I1 and CD I2, respectively, were isolated from the A. lwoffii K24 genomic library by using colony hybridization and PCR. Two DNA fragments containing the catA1 and catA2 genes were located on separate regions of the chromosome. They contained open reading frames encoding 33.4- and 30.4-kDa proteins. The amino acid sequences of the two proteins matched well with previously determined sequences. Interestingly, further analysis of the two DNA fragments revealed the locations of the catB and catC genes as well. Moreover, the DNA fragment containing catA1 had a cluster of genes in the order catB1-catC1-catA1 while the catB2-catA2-catC2 arrangement was found in the catA2 DNA fragment. These results may provide an explanation of the different substrate specificities and physicochemical properties of CD I1 and CD I2. 相似文献
63.
Multiple genes encoding 2,3-dihydroxybiphenyl 1,2-dioxygenase in the gram-positive polychlorinated biphenyl-degrading bacterium Rhodococcus erythropolis TA421, isolated from a termite ecosystem. 总被引:7,自引:3,他引:4
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Rhodococcus erythropolis TA421 was isolated from a termite ecosystem and is able to degrade a wide range of polychlorinated biphenyl (PCB) congeners. Genetic and biochemical analyses of the PCB catabolic pathway of this organism revealed that there are four different bphC genes (bphC1, bphC2, bphC3, and bphC4) which encode 2,3-dihydroxybiphenyl dioxygenases. As determined by Southern hybridization, none of the bphC genes exhibits homology to any other bphC gene. bphC1, bphC2, and bphC4 encode enzymes that have narrow substrate specificities and cleave the first aromatic ring in the meta position. In contrast, bphC3 encodes a meta cleavage dioxygenase with broad substrate specificity. Asturias et al. have shown that the closely related organism Rhodococcus globerulus P6 contains three different bphC genes (bphC1, bphC2, and bpHC3) which encode meta cleavage dioxygenases. The data suggest that there is a diverse family of bphC genes which encode PCB meta cleavage dioxygenases in members of the genus Rhodococcus. 相似文献
64.
A serotype-specific epitope of dengue virus 1 identified by phage displayed random peptide library 总被引:3,自引:0,他引:3
Zhi-Jian Yao Mandy C.C. Kao Kean-Chong Loh Maxey C.M. Chung 《FEMS microbiology letters》1995,127(1-2):93-98
Abstract From a panel of monoclonal antibodies of dengue viruses, a serotype-specific epitope of dengue virus 1 was screened from a random peptide library displayed on phage. The epitope was the determinant reactive with monoclonal antibody 15F3-1 that was specific to dengue 1. The screening was monitored by a dot blotting procedure, and after three rounds of screening a consensus motif, HRYSWK, was found. This sequence matches the sequence HKYSWK, corresponding to the amino acid residues 885–890 of polyprotein or residues 111–116 of the non-structural protein 1 of dengue virus serotype 1. The linear epitope was confirmed by testing the antigenicity of chemically synthesized 8-branched peptide. 相似文献
65.
We studied the effect of prostaglandin F2 alpha (PGF2 alpha) on the responsiveness of pulmonary airways in dogs. Airway responsiveness was assessed by determining the bronchoconstrictor response to increasing concentrations of acetylcholine aerosol delivered to the airways. In each of five dogs, we determined responsiveness during treatment with physiologic saline, histamine, or PGF2 alpha aerosols. The doses of histamine and PGF2 alpha were determined by establishing the largest dose of each which could be given to the dog without causing bronchoconstriction (subthreshold doses). We found that airway responsiveness was not significantly different during histamine treatment than after saline, however, responsiveness increased during treatment with PGF2 alpha. In addition, the hyperresponsiveness induced by PGF2 alpha was prevented by pretreatment with the ganglion blocking drug hexamethonium (5 mg/kg given intravenously). The results show that PGF2 alpha specifically increases the responsiveness of pulmonary airways in doses that do not cause bronchoconstriction, and suggest that the hyperresponsiveness involves a neural mechanism such as increased responsiveness of airway sensory nerves. 相似文献
66.
Ultrastructure and enzyme digestion of nucleoli and associated structures in hypothalamic nerve cells viewed in resinless sections 总被引:1,自引:0,他引:1
S K Chung R S Cohen D W Pfaff 《Biology of the cell / under the auspices of the European Cell Biology Organization》1984,51(1):23-33
Estrogen has been shown to affect ventromedial hypothalamic (VMH) nerve cell nucleoli in ovariectomized rats, by causing an increase in the number of electron-dense aggregates associated with nucleoli. In order to characterize these nucleolus-associated structures and other nuclear components, we examined the ultrastructure of ventromedial hypothalamic nucleoli and nuclei revealed by enzyme digestions (pepsin, RNase and DNase) in resinless thin sections. Digestion by pepsin did not cause obvious alterations in the morphology of the nucleolus or its related structures. Pepsin treatment followed by RNase, however, reduced the density of the nucleolus, while that of the nucleolus-associated structure and other related structures remained unchanged. Conversely pepsin treatment followed by DNase, reduced the density of nucleolus-associated and other chromatin structures, but had no effect on the density of the nucleolus. Pepsin treatment followed by RNase and then DNase treatment, reduced the density of the nucleolus and nucleolus-associated structures. A residual nucleolus and nucleolus-associated structure remained after this treatment. Stereo viewing of resinless sections shows that the nucleolus, its associated structures, and other related structures, are associated with fine filaments that may comprise the nuclear matrix. The nucleolus-associated structure containing DNA may direct RNA synthesis at an increased rate in estrogen-treated hypothalamic cells. 相似文献
67.
R Manjunath S I Chung A B Mukherjee 《Biochemical and biophysical research communications》1984,121(1):400-407
Uteroglobin, a progesterone induced, pregnancy related protein, can be incorporated into higher molecular weight proteins by human placental Factor XIIIa. This process is time dependent, requires CaCl2 and can be inhibited by the addition of polylysine, dansylcadavarine or histamine. Crosslinking of uteroglobin into higher molecular weight proteins can also be brought about by guinea pig liver transglutaminase. Such a process may be involved in the modification of epididymal spermatozoa to suppress their antigenicity. 相似文献
68.
Parietal yolk sac cells M1536-B3 grown on cytodex 2 beads deposited an extracellular matrix on the surface of the beads. Cell-free matrix-coated beads were isolated by treatment of the cell monolayer with cytochalasin B (CB) at a concentration of 10 μg/ml of phosphate-buffered saline (PBS). The matrix when analysed by electrophoresis on polyacrylamide gels (PAGE) revealed that the major components were laminin and entactin. The matrix-coated beads were used to study the attachment, spreading, and growth of African Green monkey BSC-40, human mammary MCF-7, mouse fibroblast L929, rat liver clone 9, and rat hepatoma H-4-II-E cells in defined serum-free growth medium. The different cell lines exhibited varying responses to matrix-coated vs uncoated beads with respect to rate of attachment, spreading, and growth. One of the most consistent responses observed was the enhancement of cell spreading on matrix-coated beads. The results suggested that the matrix-coated beads will provide a readily available and valuable tool for studies on cell surface-extracellular matrix interactions and the physiological consequences of those interactions. 相似文献
69.
Distribution of entactin in the basement membrane of the rat mammary gland. Evidence for a non-epithelial origin 总被引:4,自引:0,他引:4
M J Warburton P Monaghan S A Ferns P S Rudland N Perusinghe A E Chung 《Experimental cell research》1984,152(1):240-254
Entactin, a sulfated glycoprotein with a molecular weight (MW) of about 150 kD, is present in vascular basement membranes and in the interstitial connective tissue of the mammary glands of virgin rats. It does not appear to be present in the basement membrane surrounding the mammary ductal system. However, in lactating mammary glands entactin is also present in the basement membrane region surrounding the secretory alveoli. Ultrastructural localisation of entactin reveals that it is present on the basal surface of epithelial cells, with patchy staining in the lamina lucida and lamina densa. Entactin also appears to be associated with interstitial collagen fibres. Mammary fibroblastic cells in culture are able to produce entactin, whereas mammary epithelial and myoepithelial cells, which synthesise the basement membrane proteins laminin and type IV collagen, fail to synthesise entactin. 相似文献
70.
Rochelle S. Cohen Ph.D. Sookja Kim Chung Donald W. Pfaff 《Cell and tissue research》1984,235(3):485-489
Summary Estrogen is accumulated from the blood by nerve cells in the ventromedial nucleus of the hypothalamus and can facilitate female reproductive behavior by acting on this region of the brain. This cell group was examined in ovariectomized female rats, given estrogen or control treatment, by use of light and electron microscopy. A significantly greater portion of the nerve cells in the estrogen-treated animals had protuberances on their nucleolar surfaces, apparent under the light microscope. The fine structure of such protuberances included dense, aggregated material, which is shown to contain DNA by the sodium tungstate staining technique. Because increased numbers of such protuberances were found in nuclei of cells of the experimental group where previous studies demonstrated a significant increase in ultrastructural signs of biosynthetic activity, they may be associated with increased RNA synthesis. Thus, they could indicate, ultrastructurally, increased synthetic rates for RNA in nerve cells through which estrogen promotes reproductive behavior. 相似文献