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1.
Monoclonal antibodies raised against extracts of the rachis abscission zone of Sambucus nigra L. were selected for high reactivity towards abscission-zone proteins. One antibody (YZ1/2.23) has been shown to cross-react, by both indirect and competition enzyme-linked immunosorbent assay and by Western blotting, with a number of plant enzymes including horseradish peroxidase, rice -glucosidase, almond -glucosidase and the lectins from Phaseolus vulgaris and Erythrina cristagalli.The major N-linked oligosaccharide isolated from horseradish peroxidase has the sequence Man 3(Man6)(Xyl2)Man4GlcNAc4(Fuc3) GlcNAc. This oligosaccharide was found to be a potent inhibitor of the binding of YZ1/2.23 to the intact glycoprotein. The common determinant is therefore contained within this structure.Abbreviations ELISA enzyme-linked immunosorbent assay - Fuc fucose - GlcNAc N-acetylglucosamine - HRP horseradish peroxidase - Ig immunoglobulin - Man mannose - SDS-PAGE Sodium dodecyl sulphate-polyacrylamide gel electrophoresis - Xyl xylose  相似文献   

2.
Photoinhibition in the green alga Dunaliella salina is accompanied by the formation of inactive Photosystem II reaction centers. In SDS-PAGE analysis, the latter appear as 160 kD complexes. These complexes are structurally stable, enough to withstand re-electrophoresis of excised gel slices from the 160 kD region. Western blot analyses with specific polyclonal antibodies raised against the D1 or D2 reaction center proteins provided evidence for the presence of both of these polypeptides in the re-electrophoresed 160 kD complex. Incubation of excised gel slices from the 160 kD region, under aerobic conditions at 4°C for a prolonged period of time, caused a break-up of the 160 kD complex into a 52 kD D1-containing and 80 and 26 kD D2-containing pieces. Western blot analysis with polyclonal antibodies raised against the apoproteins of CPI (reaction center proteins of PS I) did not show cross-reaction either with the 160 kD complex or with the 52, 80 and 26 kD pieces. The results show the presence of both D1 and D2 in the 160 kD complex and strengthen the notion of a higher molecular weight D1- and D2-containing complex that forms upon disassembly of photodamaged PS II units.Abbreviations Chl chlorophyll - PS II Photosystem II - D1 the 32 kD reaction center protein of PS II, encoded by the chloroplast psbA gene - D2 the 34 kD reaction center protein of PS II, encoded by the chloroplast psbD gene - CPI the 82 and 83 kD reaction center proteins of PS I, encoded by the chloroplast psaA and psaB genes - HL high light - LL low light This publication is dedicated to the memory of the late Professor Daniel Arnon, whom the first author will fondly remember for his many accounts of past scientific discovery and debate.  相似文献   

3.
Salmon gonadotropin (GTH II) is a heterodimeric glycoprotein hormone ( and II subunits), serving as a maturational GTH, and is produced in a specific gonadotropic cell-type (GTH II-cells) containing small granules and large globules. In trout GTH II-cells, double immunolabeling for the - and II-subunits shows that colocalization of the - and II-immunolabeling is confined to the small granules, indicating storage of functional GTH II. On the other hand, -immunolabeling is absent in the large globules, even though II labeling is abundant throughout the period of seasonal gametogenesis. The -specific antiserum recognizes the intact -subunit as well as the reduced and deglycosylated -subunits by immunoblotting. These results indicate that an accumulation of the II-subunit is specifically generated in the large globules of these cells. In fact, with sexual maturity, the quantity of II-subunits becomes elevated in the trout pituitary due to a marked increase in GTH II-cells containing many large globules. However, the derivation and function of the large globules and the fate of their contained II-subunits remains unknown.  相似文献   

4.
DNA-degrading enzymes of 24.0 kDa and 27.0 kDa were observed to have different activities in two common wheat (Triticum aestivum L.) cultivars, Wichita and Cheyenne. A substrate-based SDS-PAGE assay revealed that these two enzymes were much more active in Wichita than in Cheyenne. Genes controlling the activities of these two enzymes were localized on chromosome 2D by testing DNA-degrading activities in reciprocal chromosome substitution lines between Wichita and Cheyenne. While the allele on Wichita chromosome 2D stimulated the activities of the 24.0- and 27.0-kDa enzymes in Cheyenne, the allele on Cheyenne chromosome 2D did not reduce the activities of the 24-kDa and 27-kDa enzymes in Wichita. Whether these genes code for the DNA-degrading enzymes themselves or for factors that regulate the enzyme activities remains unknown.This work was supported in part by USDA-Competitive Research Grants Office grant No. 90-37140-5426 to P.S.B. Contribution from Agricultural Research Division, University of Nebraska. Journal Series Number 10304  相似文献   

5.
Fowl plague virus, strain Dutch, was metabolically labeled withd-[2-3H]mannose, or withd-[6-3H]glucosamine, and the small subunit (HA2; 0.8 mg in total) of the viral hemagglutinin was isolated by preparative sodium dodecylsulfate-polyacrylamide gel electrophoresis. After proteolytic digestion, the radioactive oligosaccharides were sequentially liberated from the glycopeptides by treatment with different endo--N-acetylglucosaminidases and with peptide:N-glycosidase or, finally, by hydrazinolysis. In this manner, four groups of glycans could be obtained by consecutive gel filtrations and were subfractionated by HPLC. The structures of the individual oligosaccharides were analyzed by micromethylation, by acetolysis or by digestion with exoglycosidases. The major species amongst the high mannose glycans at Ans-406 of the viral glycopolypeptide were found to be Man1-2Man1-3(Man1-2Man1-6)Man1-6(Man1-2Man1-2Man1-3)Man1-4GlcNac1-4GlcNAc and Man1-3(Man1-2Man1-6)Man1-6(Man1-2Man1-2Man1-3)Man1-4GlcNAc1-4GlcNAc, while the complex glycans at Asn-478 are predominantly GlcNAc1-2Man1-3(GlcNAc1-2Man1-6)Man1-4GlcNAc1-4GlcNAc (lacking, in part, one of the outerN-acetylglucosamine residues) and GlcNAc1-2Man1-3(Gal1-4GlcNAc1-2Man1-6)Man1-4GlcNAc1-4GlcNAc.Abbreviation BSA bovine serum albumin - endo D (F,H) endo--N-acetyl-d-glucosaminidase D (F,H) - HA hemagglutinin (HA1, large subunit of HA - HA2 small subunit - FPV fowl plague virus - PNGase F peptide:N-glycosidase F - SDS sodium dodecylsulfate  相似文献   

6.
The brain and thoracic ganglion neurosecretory cells of the freshwater prawn, M. kistnensis displayed characteristic changes following 24 hr exposure to warm (31.0° ± 0.5°C) and cold (13.0° ±0.5°C) stress. A and B cells from the brain showed significant elevation in their nuclear diameters after warm and cold stress. Warm stress depressed whereas cold stress enhanced the neurosecretory material intensity of A cells. However, the intensity of B cells neurosecretory material remained unaltered following both treatments. On the contrary nuclear diameters of both A and B neurosecretory cell significantly decreased and an increment in neurosecretory material intensity was revealed in the thoracic ganglion of both warm and cold stressed prawns.  相似文献   

7.
Rabbit antisera were raised against -(16)-galactotetraose coupled to bovine serum albumin (Gal4-BSA). The antisera reacted with arabinogalactan-proteins (AGPs) isolated from seeds, roots, or leaves of radish (Raphanus sativus L.) as revealed by immunodiffusion analysis. Extensive removal of -l-arabinofuranosyl residues from these AGPs enhanced the formation of precipitin with the antisera. The antisera did not react with such other polysaccharides as soybean arabinan-4-galactan, -(14)-galactan, and -(13)-galactan, indicating their high specificity toward the consecutive -(16)-galactosyl side chains of AGPs. The antibodies were purified by affinity chromatography on a column of immobilized -(16)-galactotetraose as ligand. The specificity of the antibodies toward consecutive (16)-linked -galactosyl residues was confirmed by enzyme-linked immunosorbent assay for hapten inhibition against Gal4-BSA as antigen, which revealed that -(16)-galactotriose and-tetraose were potent inhibitors, while -(13)-or -(14)-galactobioses and -trioses were essentially unreactive. Electron-microscopic observation of immunogold-stained tissues demonstrated that AGPs were localized in the middle lamella as well as at the plasma membrane of primary roots of radish. Agglutination of protoplasts prepared from cotyledons occurred with the antibodies, supporting the evidence for localization of AGPs in the plasma membrane. The antibody-mediated agglutination was inhibited by addition of AGPs or -(16)-galactotetraose.Abbreviations AGP arabinogalactan-protein - BSA bovine serum albumin - ELISA enzyme-linked immunosorbent assay - FITC fluorescein isothiocyanate - Gal3-BSA -(16)-galactotriose coupled to BSA - Gal4-BSA -(16)-galactotetraose coupled to BSA - Ig immunoglobulin - 4-Me-GlcpA 4-O-methyl-d-glucopyranosyluronic acid - Mr relative molecular mass The authors wish to thank Dr. J. Ohnishi of Department of Biochemistry, Saitama University, for his help in preparing protoplasts.  相似文献   

8.
Functionally active preparations of Na+,K+-ATPase isozymes from calf brain that contain catalytic subunits of three types (1, 2, and 3) were obtained using two approaches: a selective removal of contaminating proteins by the Jorgensen method and a selective solubilization of the enzyme with subsequent reconstitution of their membrane structure by the Esmann method. The ouabain inhibition constants were determined for the isozymes. The real isozyme composition of the Na+ pump from the grey matter containing glial cells and the brain stem containing neurons was determined. The plasma membranes of glial cells were shown to contain mainly Na+,K+-ATPase of the 11 type and minor amounts of isozymes of the 22(1) and the 31(2) type. The axolemma contains 21 and 31 isozymes. A carbohydrate analysis indicated that 11 enzyme preparations from the brain grey matter substantially differ from the renal enzymes of the same composition in the glycosylation of the 1 isoform. An enhanced sensitivity of the 3 catalytic subunit of Na+,K+-ATPase from neurons to endogenous proteolysis was found. A point of specific proteolysis in the amino acid sequence PNDNR492 Y493 was localized (residue numbering is that of the human 3 subunit). This sequence corresponds to one of the regions of the greatest variability in 1-, 2-, 3-, and 4-subunits, but at the same time, it is characteristic of the 3 isoforms of various species. The presence of the 3 isoform of tubulin (cytoskeletal protein) was found for the first time in the high-molecular-mass Na+,K+-ATPase 31 isozyme complex isolated from the axolemma of brain stem neurons, and its binding to the 3 catalytic subunit was shown.  相似文献   

9.
K. J. Föhr  M. Enßle  H. Schraudolf 《Planta》1987,171(1):127-129
Spores and prothallia of the fern Anemia phyllitidis L. Sw. contain a protein which in its physicochemical properties corresponds largely to calmodulin. It shows immunoreactivity with a calmodulin antiserum and activates bovine brain phosphodiesterase. Its content increases during the early processes of light-induced spore germination, indicating that the Ca2+-dependence of these processes may be mediated by this protein.Abbreviations EGTA ethylene, glycol-bis(-aminoethyl ether)-N,N,N,N-tetraacetic acid - RIA radioimmunoassay  相似文献   

10.
Summary Tobacco plants (Nicotiana tabacum L.) of four varieties (Badischer Burley, White Burley, Techne, Kupchunos) were raised at different temperatures and daylengths and the effect of genotype on embryogenic pollen grain formation in situ and on pollen plant formation in anther and pollen cultures from these plants was studied. Genotype controlled embryogenic pollen grain and pollen plant formation by defining productivity under standard growth conditions (long days at 24 °C). Kupchunos was the most productive variety, followed by White Burley, Techne, and Badischer Burley. Furthermore, genotype defined which environmental factor was able to affect embryogenic pollen grain and pollen plant formation and also to which degree. In anther cultures, in addition to these effects, genotype controlled the formation of (an) inhibitory substance(s) in the anther wall in interaction with the plant growth conditions. In Badischer Burley and Techne, inhibitor action could be prevented by isolation of the pollen after one week of anther culture. Finally, direct pollen cultures in Badischer Burley and Techne produced embryos were only when the pollen was isolated from nearly mature anthers, while in White Burley and Kupchunos, embryos also produced at earlier stages and at higher yields. This indicated that genotype controls the time when the embryogenic pollen grains become ready to divide. The results are discussed in relation to strategies to overcome recalcitrance of species and genotypes.  相似文献   

11.
I investigated the influence of insemination at different life stages on female fitness in the mite Histiostoma feroniarum. In this species, males guard immature females at the tritonymph stage to inseminate them immediately after the last moulting. Four groups of females were studied (1) females inseminated naturally, i.e. mating occurred immediately after guarding, and then the male was removed (IF/0M), (2) naturally inseminated females, where after insemination the male was replaced by two additional males (IF/2M), (3) virgin females reared without males (VF/0M) and (4) mature, virgin females to which two virgin males were added 3 days after last moulting (VF/2DM). The results show that females inseminated naturally (IF/0M) have higher longevity and fecundity than either virgin (VF/0M) or late-inseminated females (VF/2DM). Furthermore, longevity and fecundity of the former (IF/0M) was also greater than that of females naturally inseminated and subsequently exposed to males (IF/2M). One may suggest that seminal fluids have a positive effect on female fitness. When delayed insemination occurs, such positive effect may not be observed due to a change in features of the sperm access system. Harassment may explain decreased longevity and fecundity of females inseminated naturally compared to females that received additional males.This revised version was published online in May 2005 with a corrected cover date.  相似文献   

12.
A linkage map with RFLP and isozyme markers for almond   总被引:12,自引:0,他引:12  
Inheritance and linkage studies were conducted with seven isozyme genes and 120 RFLPs in the F1 progeny of a cross between almond cultivars Ferragnes and Tuono. RFLPs were detected using 57 genomic and 43 cDNA almond clones. Eight of the cDNA probes corresponded to known genes (extensin, prunin (2), -tubulin, endopolygalacturonase, oleosin, actin depolymerizing factor and phosphoglyceromutase). Single-copy clones were found more frequently in the cDNA (65%) than in the genomic libraries (26%). Two maps were elaborated, one with the 93 loci heterozygous in Ferragnes and another with the 69 loci heterozygous in Tuono. Thirty-five loci were heterozygous in both parents and were used as bridges between both maps. Most of the segregations (91%) were of the 11 or 1111 types, and data were analyzed as if they derived from two backcross populations. Eight linkage groups covering 393 cM in Ferragnes and 394 in Tuono were found for each map. None of the loci examined in either map was found to be unlinked. Distorted segregation ratios were mainly concentrated in two linkage groups of the Ferragnes map.  相似文献   

13.
Morniga M is a jacalin-related and mannose-specific lectin isolated from the bark of the mulberry (Morus nigra). In order to understand the function and application of this novel lectin, the binding property of Morniga M was studied in detail using an enzyme-linked lectinosorbent assay and lectin-glycan inhibition assay with extended glycan/ligand collection. From the results, it was found that the di-, tri-, and oligomannosyl structural units of N-glycans such as those of the bovine 1-acid glycoprotein (gp) and lactoferrin were the most active gps, but not the O-glycans or polysaccharides including mannan from yeast. The binding affinity of Morniga M for ligands can be ranked in decreasing order as follows: gps carrying multiple N-glycans with oligomannosyl residues >> N-glycopeptide with a single trimannosyl core > Tri-Man oligomer [Man1 6(Man 1 3) Man], Penta-Man oligomer [Man1 6(Man1 3)Man1 6(Man1 3) Man] Man 1 2, 3 or 6 Man > Man > GlcNAc, Glc >> L-Fuc, Gal, GalNAc (inactive), demonstrating the unique specificity of this lectin that may not only assist in our understanding of cell surface carbohydrate ligand-lectin recognition, but also provide informative guidelines for the application of this structural probe in biotechnological and clinical regimens, especially in the detection and purification of N-linked glycans.  相似文献   

14.
Crude extracts or supernatants of broken cells of Clostridium formicoaceticum reduce unbranched, branched, saturated and unsaturated carboxylates at the expense of carbon monoxide to the corresponding alcohols. The presence of viologens with redox potentials varying from E 0=-295 to-650 mV decreased the rate of propionate reduction. The more the propionate reduction was diminished the more formate was formed from carbon monoxide. The lowest propionate reduction and highest formate formation was observed with methylviologen. The carbon-carbon double bond of E-2-methyl-butenoate was only hydrogenated when a viologen was present. Formate as electron donor led only in the presence of viologens to the formation of propanol from propionate. The reduction of propionate at the expense of a reduced viologen can be followed in cuvettes. With respect to propionate Michaelis Menten behavior was observed. Experiments are described which lead to the assumption that the carboxylates are reduced in a non-activated form. That would be new type of biological reduction.Non-standard abbreviations glc Gas liquid chromatography - HPLC high performance liquid chromatography - RP reverse phase; Mediators (the figures in parenthesis of the mediators are redox potentials E 0 in mV) - CAV2+ carbamoylmethylviologen, 1,1-carbamoyl-4,4-dipyridinium dication (E 0=-296 mV) - BV2+ benzylviologen, 1,1-dibenzyl-4,4-dipyridinium dication (E 0=-360 mV) - MV methylviologen, 1,1-dimethyl-4,4-dipyridinium-dication (E 0=-444 mV) - DMDQ2+ dimethyldiquat, 4,4-dimethyl-2,2-dipyridino-1,1-ethylendication (E 0=-514 mV) - TMV2+ tetramethylviologen, 1,1,4,4-tetramethyl-4,4-dipyridinium dication (E 0=-550 mV) - PDQ2+ propyldiquat, 2,2-dipyridino-1,1-propenyl dication (E 0=-550 mV) - DMPDQ2+ dimethylpropyldiquat, 4,4-dimethyl-2,2-dipyridino-1,1-propenyl dication (E 0=-656 mV) - PN productivity number=mmol product (obtained by the uptake of one pair of electrons) x (biocatalyst (dry weight) kg)-1×h-1  相似文献   

15.
Bycatch: from emotion to effective natural resource management   总被引:1,自引:0,他引:1  
Although bycatch has been an acknowledged component of fishery management for many years, its explosive growth as a major management issue has occurred over the past decade. Emergence of the bycatch issue can be traced to increasing world competition for the ocean's fishery resources and the rapid rise of the conservation and environmental movement in recent years. Bycatch has in the past several years been equated to waste, non-selective fishing methods that threaten non-target species and degradation of ocean ecosystems. Although some important steps have been taken to abate bycatch, frequently the problem is not perceived as a component of fishery management, e.g. establishing bycatch mortality rates and evaluating their consequences upon affected populations. Fishery scientists often estimate levels of bycatch, but the overall consequences of discard mortality resulting from the complex of fisheries operating in a region are frequently unknown and speculative. The sum of fishery-induced mortalities occurring as a result of harvesting often involves a significant number of fish in addition to catch and discard. An ICES study group has characterized fishing mortality as the aggregate of all catch mortalities including discard, illegal fishing and misreporting. It is unlikely that managers will, in the near future, have a full accounting of unobserved fishing mortality. Progress toward identifying and measuring such mortalities is receiving increasing attention.The authors suggest and provide an example of a matrix-type analysis for recording bycatch and other fishing mortalities. The matrix presentation would allow managers to evaluate various fishery-related mortality factors and their importance in developing management strategies.  相似文献   

16.
Summary The inheritance of the dwarf plant type was studied in blackgram (V. mungo (L.) Hepper). Type 9 has erect plant type with normal internode length. The mutant line, EMSD has reduced internode length. The F1, F2 and F3 generations of a cross between Type 9 and EMSD and its reciprocal were studied. The extreme dwarf plant type appeared to be governed by a single recessive gene, dw 1 dw 1 with no cytoplasmic effect.Part of Ph.D. Thesis submitted by the first author  相似文献   

17.
Beans (Phaseolus vulgaris) are regarded as a susceptible crop to suboptimal temperatures. In temperate regions, low temperatures reduce establishment of beans when planted early in the growing season. Seeds of 14 cultivars/lines or beans were germinated in petri dishes at a constant 8, 10, 12, or 18°C or at 12 h alternating temperatures of 10/8, 12/8 or 18/8°C. Differences in germination percentages and rates between cultivars/lines were significant, especially at low temperatures. Cultivars/lines that germinated best and quickly at constant 8°C were Volare, Great Northern (G.N.) Tara, G.N. Belneb # 1, G.N. Spinel, and San Cristobal. Germination percent and rate of Pinto-UI-111 and Canadian Wonder increased significantly when temperatures were increased by 2 to 4°C for 12 h per 24 h, compared with a constant 8°C. Whereas, germination of G.N. Belneb # 1 was reduced. Polyacrylamide gel electrophoresis was used to study the effect of cold treatment on polypeptide patterns of seven cultivars/lines. Seeds were germinated at 18°C constant for 96 h or at 18°C for 48 h followed by 48 h at 2 or 8°C. During cold treatment the synthesis of some polypeptides increased. Volare, G.N. Tara Pinto-UI-111 and Canadian Wonder showed changes in polypeptide patterns, while Alubia-33-1, Michigan 84100 and BAT-1225 showed no changes in polypeptide patterns if compared to the control (96 h at 18°C in the dark). This suggests a likely essential role of these proteins in the development of chilling tolerance.  相似文献   

18.
Some components of natural foods may enhance or inhibit prostaglandin formation and potentially affect the inflammation condition. A macrophage cell line, RAW264.7, was employed to examine the effects of foods traditionally regarded as heating or cooling on the production of PGE2, a well-known proinflammatory mediator. Foods traditionally regarded as heating (litchi, longan, and dried longan) or cooling (chrysanthemum flower, bitter gourd, and lotus seed plumule) were extracted sequentially with water and ethyl acetate. The water extracts (WE) and ethyl acetate extracts (EAE) were applied to RAW264.7 macrophages in the presence or absence of LPS (lipopolysaccharide). In the absence of LPS, the WEs from the heating foods, litchi, longan, or dried longan had a dose-dependent enhancing effect on PGE2 production, with respective EC50s of 8.4, 16, and 11 mg/ml. This effect was accompanied by significant induction of COX-2 protein expression, as shown by Western blot analysis. In contrast, LPS-induced PGE2 production was inhibited in a dose-dependent manner by the WEs of the cooling foods, chrysanthemum flower, bitter gourd, and lotus seed plumule, with respective IC50s of 0.6, 0.13, and 0.08 mg/ml. At the concentrations tested, none of the EAEs had any effect on basal PGE2 production, while LPS-induced PGE2 production was inhibited or increased by the EAE from bitter gourd and longan, respectively. Water-soluble extracts of foods traditionally regarded as heating enhanced basal PGE2 production, while those from cooling foods significantly inhibited LPS-induced PGE2 production by the macrophage cell line. This subject merits further study to determine whether appropriate food selection may help patients suffering from chronic inflammatory conditions.  相似文献   

19.
Sensitive immunologic techniques are now available for the detection and quantitation of carcinogen-DNA adducts. We have developed a number of specific monoclonal antibodies which recognize DNA modified by particular carcinogens, including benzo[a]-py-rene, aminopyrene, 8-methoxypsoralen plus UVA light and vinyl chloride. These antibodies can be used in competitive enzyme-linked immunosorbent assays to quantitate adducts in DNA isolated from biological samples. Samples from treated animals as well as from humans with occupational or environmental exposure to carcinogens have been studied. In addition, antibodies can be used in indirect immunohistochemical studies to localize adduct formation in various tissues or cell types.Abbreviations BPDE-1 7,8-dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene - CTCL Cutaneous T cell lymphoma - ELISA enzyme linked immunosorbent assay - 8-MOP 8-methoxypsoralen - PUVA 8-MOP plus UVA irradiation  相似文献   

20.
Summary High frequencies of callusing were obtained in indica rice from the microspores which were staged in acetic acid iron alum-haematoxylin stain prior to culture on G5 medium. Two local varieties, Khonorullo and Namyi, and two advance pre-release cultures, PK 1-1-3 and PK 12-22, were used in this investigation. All the cultures exhibited a wide adaptation to varying medium; however, the frequency of callusing was highest (45.5%) in PK 1-1-3 followed by PK 12-22 (32.4%) and Khonorullo (31.6%). Cold shock (10 °C) for 11 days enhanced the frequency of callusing by 200% in Khonorullo.  相似文献   

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