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41.
Characteristics and epitope mapping of a cloned human autoantigen La 总被引:13,自引:0,他引:13
A D Sturgess M G Peterson L J McNeilage S Whittingham R L Coppel 《Journal of immunology (Baltimore, Md. : 1950)》1988,140(9):3212-3218
The La (SS-B) polypeptide is a ribonucleoprotein against which high titer antinuclear antibodies (ANA) react in the human autoimmune disease primary Sj?gren's syndrome. To identify the autoepitopes with which the ANA anti-La (anti-SS-B) reacts, we isolated a 1.4-kb cDNA clone for La from a lambda gt10 library made from a human Burkitt's cell line. This clone contained an open reading frame of 1065 bp, encoding a 40.1-kDa polypeptide that corresponded to the carboxyl-terminal end of the La protein. The predicted polypeptide sequence of the recombinant protein was highly charged and unrelated to any previously published sequence. We also compared this clone to a previously published cDNA sequence for La and demonstrated significant differences, particularly that the open reading frame in our cDNA continued for 926 additional bases 3' to a putative termination codon in the previously reported sequence. The recombinant La protein was expressed in Escherichia coli and tested for reactivity with 200 sera containing ANA of various specificities. Only the sera containing anti-La antibodies reacted with the cloned La. By expressing subclones of the La cDNA as fusion proteins with beta-galactosidase, we have localized at least one epitope for the binding of anti-La antibodies to the carboxyl-terminal 103 amino acids of the La protein. No anti-La binding could be demonstrated to the region of the La protein that had previously been predicted to contain an autoepitope for the binding of anti-La (SS-B) antibodies. Studies of cloned autoepitopes could provide important clues to the role ANA play in disease and lead to targeted intervention in the treatment of primary Sj?gren's syndrome. 相似文献
42.
Summary The relative hydraulic conductivities of major and minor longitudinal veins, and the apoplastic permeability of the bundle sheaths surrounding all longitudinal and transverse veins were investigated in representatives of the C3, C4/NAD-ME, C4/NAD-ME/PCK intermediate, C4/PCK and C4/NADP-ME photosynthetic types. Using the Hagen-Poiseuille equation and measurements of tracheary element diameters, the number of elements in each vein type and the numbers of each vein type, we calculated that 87–99% of the water flow in a longitudinal direction would be expected to occur in the major veins. The permeability of the mestome sheaths and parenchymatous bundle sheaths surrounding the veins was tested using the negatively-charged, fluorescent dye, trisodium 3-hydroxy-5,8,10-pyrenetrisulfonate (PTS). This dye proved nontoxic to plant tissue at a concentration of 0.5%, according to a deplasmolysis test with onion epidermal strips. The PTS concentration achieved in the tested grass leaves was about 0.035%, well below the toxic limit. When a solution of PTS was fed to the leaves by means of a basal cut, the dye moved into the veins of all orders. From there, it moved outward into the surrounding tissues, indicating that the sheaths surrounding the veins of all orders in all species tested were permeable. Therefore, contrary to previous predictions based on structural observations and some tracer studies, bundle sheaths with suberized cell walls do not function as endodermal layers. 相似文献
43.
A berberine-aniline blue fluorescent staining procedure for suberin,lignin, and callose in plant tissue 总被引:17,自引:0,他引:17
Summary A fluorescent staining procedure to detect suberin, lignin and callose in plants has been developed. This procedure greatly improves on previous methods for visualizing Casparian bands in root exodermal and endodermal cells, and performs equally well on a variety of other plant tissues. Berberine was selected as the most suitable replacement forChelidonium majus root extract after comparing the staining properties of the extract with those of four of its constituent alkaloids. Aniline blue counterstaining efficiently quenched unwanted background fluorescence and nonspecific berberine staining, while providing a fluorochrome for callose. When used with multichambered holders which allow simultaneous processing of freehand sections, this efficient staining procedure facilitates morphological studies involving large numbers of samples.Abbreviations ISCC-NBS
Inter-Society Color Council-National Bureau of Standards
- UV
ultraviolet light 相似文献
44.
Simultaneous determination of ascorbic acid and dehydroascorbic acid in cultures of C3H/10T1/2 cells
Luminita L. V. Ibric William F. Benedict Andrew R. Peterson 《In vitro cellular & developmental biology. Plant》1988,24(7):669-676
Summary A reproducible method is described for the separation and quantification of ascorbic acid and dehydroascorbic acid by ion-pairing
reverse-phase high performance liquid chromatography and detection by absorbance at 232 nm. Lowest detectable concentrations
with a linear response of detection were 5 nmol for ascorbic acid and 50 nmol for dehydroascorbic acid. This method was applied
to the analysis of C3H/10T1/2 cells and culture medium after influx or efflux experiments and single or multiple treatments
with ascorbic acid. Subsequent measurement of the radioactivity in the eluted fractions increased the detectability of both
ascorbic acid and dehydroascorbic acid to 10 to 20 pmol.
This research was supported by grant CA 09320 and CA 31574 from the National Cancer Institute, Bethesda, MD, and grant BC441
from The American Cancer Society. 相似文献
45.
Agrobacterium tumefaciens is established as a vector for gene transfer in many dicotyledonous plants but is not accepted as a vector in monocotyledonous plants, especially in the important Gramineae. The use of Agrobacterium to transfer genes into monocot species could simplify the transformation and improvement of important crop plants. In this report we describe the use of Agrobacterium to transfer a gene into corn, the regeneration of plants, and detection of the transferred genes in the F1 progeny. Shoot apices of Zea mays L. variety Funk's G90 were cocultivated with A. tumefaciens EHA 1, which harbored the plasmid pGUS3 containing genes for kanamycin resistance (NPT II) and β-glucuronidase (GUS). Plants developed from these explants within 4 to 6 weeks. Fluorometric GUS assays of leaves and immature seeds from the plants exhibited low GUS activity. Both NOS and GUS gene fragments were amplified by polymerase chain reaction in the DNA isolated from the F1 generations of one of the original transformed plants. Southern analysis showed both GUS and NPT probes hybridized to DNA in several of the F1 progeny, demonstrating the incorporation of GUS and NPT II genes into high molecular weight DNA. These data establish successful gene transfer and sexual inheritance of the genes. 相似文献
46.
Effects of O(2) and CO(2) Concentrations on Quantum Yields of Photosystems I and II in Tobacco Leaf Tissue 总被引:3,自引:3,他引:0
Peterson RB 《Plant physiology》1991,97(4):1388-1394
The interactive effects of irradiance and O2 and CO2 levels on the quantum yields of photosystems I and II have been studied under steady-state conditions at 25°C in leaf tissue of tobacco (Nicotiana tabacum). Assessment of radiant energy utilization in photosystem II was based on changes in chlorophyll fluorescence yield excited by a weak measuring beam of modulated red light. Independent estimates of photosystem I quantum yield were based on the light-dark in vivo absorbance change at 830 nanometers, the absorption band of P700+. Normal (i.e. 20.5%, v/v) levels of O2 generally enhanced photosystem II quantum yield relative to that measured under 1.6% O2 as the irradiance approached saturation. Photorespiration is suspected to mediate such positive effects of O2 through increases in the availability of CO2 and recycling of orthophosphate. Conversely, at low intercellular CO2 concentrations, 41.2% O2 was associated with lower photosystem II quantum yield compared with that observed at 20.5% O2. Inhibitory effects of 41.2% O2 may occur in response to negative feedback on photosystem II arising from a build-up in the thylakoid proton gradient during electron transport to O2. Covariation between quantum yields of photosystems I and II was not affected by concentrations of either O2 or CO2. The dependence of quantum yield of electron transport to CO2 measured by gas exchange upon photosystem II quantum yield as determined by fluorescence was unaffected by CO2 concentration. 相似文献
47.
Summary The ectomycorrhizal fungus,Paxillus involutus, produces sclerotia in culture. These can be induced to form on agar medium by exposing mycelium grown at 25°C to various temperatures between6°C and 15°C. Sclerotia formed at 10°C and above were large and covered with drops of exudate, while those formed at 6°C or 8°C were very small and did not produce an exudate. Mature sclerotia were bounded by a compact rind and contained abundant storage reserves. Histochemistry of the larger sclerotia showed large quantities of protein stored as protein bodies in the cytoplasm, lipid present as small droplets, glycogen granules stored in the cytoplasm and polyphosphate present as small granules in the cytoplasm and in the protein bodies. Energy dispersive X-ray microanalysis confirmed the presence of phosphate in the granules and was used to map its distribution throughout the sclerotium. The smaller sclerotia induced at 8°C and below on the same medium had the same basic structure and composition, but lacked the complex phenolic cell network found in large sclerotia, and had abundant extracellular polysaccharides. The rind was not well developed and these small sclerotia are interpreted to have been arrested at an early stage of development. 相似文献
48.
Structural aspects of Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco] tuberculate ectomycorrhizae
H. B. Massicotte L. H. Melville C. Y. Li R. L. Peterson 《Trees - Structure and Function》1992,6(3):137-146
Summary Tubercles of Pseudotsuga menziesii consisted of clusters of ectomycorrhizae surrounded by a peridiumlike rind. Energy dispersive spectroscopy demonstrated that crystals found in the zone of loose hyphae extending from the inner rind to the mantle of each root probably contain calcium oxalate. Inner mantle and Hartig net hyphae showed a labyrinthine branching pattern and stored carbohydrates and protein. The Hartig net formed up to inner cortical cells which had thickened, darkly stained walls. Bacteria were located either along with hyphae within the rind or as colonies on the surface of the tubercle. 相似文献
49.
MHC class II molecules present internalized antigens to the immune system. They have long been known to associate with a polypeptide called the invariant chain. Recent findings have revealed that this polypeptide performs two functions. First, it prevents class II molecules from binding antigenic peptides at the site of synthesis of class II molecules in the endoplasmic reticulum.Second, it targets class II molecules to their destination in the endocytic pathway, where they pick up antigenic peptides derived from endocytosed antigens. Short sequences in the cytoplasmic portion of the invariant chain serve as subcellular address labels.The functions of the invariant chain help to explain how the immune system divides its defence against foreign pathogens between cytotoxic T cells and antibodies. 相似文献
50.