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91.
Catherine Digonnet-Kerhoas Gilles Gay Jean Claude Duplan Christian Dumas 《Planta》1989,179(2):165-170
During ageing of the short-lived pollen grains of Cucurbita pepo L., water loss was examined in relation to viability using biophysical (1H-nuclear magnetic resonance, NMR) and cytological methods (fluorochromatic reaction test, freezefracture and scanning electron microscopy). A semi-logarithmic representation of the pollen weight loss demonstrated the complexity of the dehydration process. A the study of proton loss using 1H-NMR indicated that two major releases water of had taken place, each with different flux rates. Pulse 1H-NMR experiments showed the occurrene of non-exponential signal decay as a function of time, indicating the existence of different fractions of water in a pollen grain sample. These fractions leave the pollen grain at different times during pollen dehydration, and one of them (that of the so-called vital water) can be related to pollen viability. The quantity of protons giving a signal during pulse 1H-NMR experiments was very low when the pollen grains were judged to be dead according to the fluorochromatic test. Freeze-fracture replicas of these dead pollen grains (less than 25% water content) showed that the plasma membrane had become detached from the intine surface; this ultrastructural feature might therefore be involved in the loss of pollen viability.Abbreviations A
initial amplitude of the NMR signal
- A2
quantity of water charcterized by T2-2
- A5
quantity of water characterized by T2–5
- FCR
fluorochromatic reaction
- NMR
nuclear magnetic resonance
- T2
transverse relaxation time
- T2-2
T2 measured with 2 ms between each pulse of radiofrequency
- T2–5
T2 measured with 5 ms between each pulse of radiofrequency 相似文献
92.
In Bolivia, the dog is involved in the cycle of visceral leishmaniasis (Leishmania (Le.) chagasi) in the Yungas (alt. 1,000-2,000 m), and also in the cycle of cutaneous leishmaniasis (Le. (V.) braziliensis) in the Alto Beni (alt. 400-600 m). But it plays a different role in the two cycles. In the Yungas focus, it is the main reservoir of Le. (Le.) chagasi and the source of contamination for man. In the Alto Beni focus, it is only a "victim-host", like man, of Le. (V.) braziliensis; the reservoir of which is unknown. Wild mammals are very likely to be involved. 相似文献
93.
Ossarath Kol Colette Brassart Geneviéve Spik Jean Montreuil Stéphane Bouquelet 《Glycoconjugate journal》1989,6(3):333-348
We have previously shown that an endo--N-acetylglucosaminidase (EC 3.2.1.96) named Endo B, isolated from culture filtrates of the basidiomyceteSporotrichum dimorphosporum cleaves asialo-, and to some extent, monosialylated bi-antennary glycans of theN-acetyllactosamine type linked to the asparagine residue of peptide or protein moieties [Bouquelet S, Strecker G, Montreuil J, Spik G (1980) Biochimie 62:43–49]. In the present paper, the substrate specificity of the enzyme towards oligomannoside and hybrid type glycans has been analyzed. The results obtained indicate that ovalbumin glycopeptides containing four to seven mannose residues and bovine lactotransferrin glycopeptides containing four to nine mannose residues were completely hydrolyzed by the enzyme. The degree of cleavage was variable among hybrid type structures, since glycopeptides containing the following glycans: (Gal)1(GlcNAc)3(Man)5(GlcNAc)2; (GlcNAc)3(Man)5(GlcNAc)2; (GlcNAc)3(Man)4(GlcNAc)2 were not hydrolyzed by the enzyme while the percentage of hydrolysis of a glycopeptide containing (GlcNAc)2(Man)5(GlcNAc)2 glycan reached 90%. The bovine lactotransferrin was partially deglycosylated (40%) in the absence of non-ionic detergent while native ovalbumin glycoprotein was not hydrolyzed by the enzyme.The oligomannoside-and theN-acetyllactosamine-type degrading activities present in the culture filtrates were not separated at any step of the purification procedure. Both activities were eluted as a single component with an apparent molecular mass of 89 kDa suggesting that they are located on the same enzyme molecule.Endo B represents a powerful tool for removing oligomannoside-andN-acetyllactosamine-type glycans fromN-glycopeptides andN-glycoproteins. Moreover, advantages in the use of Endo B in a soluble form as well as in an immobilized form result in its high activity and in its stability to heat denaturation and storage.Abbreviations Gal
d-galactose
- Man
d-mannose
- GlcNAc
N-acetyl-d-glucosamine
- Con A
concanavalin A
- Asn
asparagine
- GLC
gas liquid chromatography
- TLC
thin layer chromatography
- Endo
endo--N-acetylglucosaminidase
- Endo B
endo--N-acetylglucosaminidase isolated fromSporotrichum dimorphosporum
- PBE
polybuffer exchanger
- SDS-PAGE
sodium dodecylsulfate-polyacrylamide gel electrophoresis 相似文献
94.
Tobacco cells are sensitive to bleomycin and phleomycin. The Tn5 and the Streptoalloteichus hindustanus (Sh) bleomycin resistance (Ble) genes conferring resistance to these antibiotics have each been inserted into two plant expression vectors. They are flanked by the nopaline synthase (nos) or the cauliflower mosaic virus (CaMV) 35S promoters on one side, and by the nos polyadenylation signal on the other. These four chimaeric genes were introduced into the binary transformation vector pGA 492, which were thereafter mobilized into Agrobacterium tumefaciens strain LBA 4404. The resulting strains were used to transform Nicotiana tabacum cv. Xanthi nc using the leaf disc transformation procedure. In all cases, phleomycin- and bleomycin-resistant tobacco plants were regenerated from transformed cells under selective conditions; however the highest frequency of rooted plants was obtained when transformation was carried out with the Sh Ble gene under the control of the 35S promoter. Phleomycin resistance was stably transmitted to sexual offspring as a dominant nuclear trait as confirmed by Southern blotting. 相似文献
95.
Catherine Manin Jean Noël Barbotin Daniel Thomas Jean Claude Lazzaroni 《Applied microbiology and biotechnology》1989,32(2):143-147
Summary In continuous cultures, alkaline phosphatase was synthesised and excreted for more than 250 h by immobilized growing cells in contrast to free cells for which the excretion decreased after 150 h of culture. This observed increase in alkaline phosphatase synthesis and excretion by immobilized cells may have resulted from growing conditions within the gel beads.Offprint requests to: C. Manin 相似文献
96.
Slim-Eric Letaíef Grard Landemore Jean Bocquet Jacques Izard 《Biology of the cell / under the auspices of the European Cell Biology Organization》1989,65(3):257-263
This paper reports that the Kurloff cell sulphated and chondroitinase AC sensitive material previously described filtered on Sepharose CL4B columns as 2 main populations with Kav of 0.25 and 0.44. Its alkaline treatment resulted in the elution of 2 peaks with Kav of 0.52 and 0.78. Their reduction in size observed after alkaline treatment and the 6-fold increase in the (35S) sulphate incorporation after addition of 0.1 mM xyloside to the incubation medium indicate that these intracellular sulphated glycosaminoglycans exist in the form of proteoglycans. They were characterized by their resistance to degradation by pronase, papain or cathepsin D, as assessed by gel filtration chromatography on Sepharose CL6B or CL4B. After the glycosaminoglycans were digested with chondroitinase AC, thin-layer chromatography analysis indicated the presence of delta di-4S and delta di-6S in a ratio of 7:1. The presence of such protease-resistant proteochondroitin sulphate in intracytoplasmic granules of both Kurloff cells and other natural killer cell types is emphasized. 相似文献
97.
Sequence divergence and open regions of RNA secondary structures in the envelope regions of the 17 human immunodeficiency virus isolates. 总被引:4,自引:2,他引:2 下载免费PDF全文
Genetic variation during the course of infection of an individual is a remarkable feature of the acquired immune deficiency syndrome (AIDS) disease. This variation has been studied for the envelope protein encoding regions of seventeen different sequences from various isolates of human immunodeficiency virus (HIV) using multiple sequence comparison and calculation of variability. The open regions with little intramolecular base pairing in these envelope sequences are predicted by a recently developed statistical method. The minimum length L for a run of hypervariable sites, conserved sites, or open regions that gives significance at the 1% (or 0.1%) level is then determined by a scan statistical method. The results show that significant clusters of open regions predicted at the RNA levels correlate with significant clusters of hypervariable sites in the HIV envelope gene. Those significant genomic variations in HIVs seem to be manifested mainly in the extracellular portion of the envelope protein. Twelve potential antigenic determinants are predicted using an antigenic index method. Interestingly, the majority of the significant hypervariable regions in the exterior envelope protein (gp120) were predicted potential epitopes. 相似文献
98.
Purification and properties of the DNA-binding protein HPB12 from Bacillus subtilis nucleoid 总被引:2,自引:0,他引:2
We report the purification to homogeneity of a 12 KDa protein (HPB12) present in the nucleoids of Bacillus subtilis. From the purification data the abundance of the protein was estimated to about 20,000 monomers per cell. The HPB12 protein is heat-stable and acid-soluble and binds preferentially to supercoiled and linearized double-stranded DNAs. The binding of the protein to the supercoiled DNA occurs very rapidly and appears to be cooperative. Moreover, the complexes are extremely stable and do not dissociate after 90 min. These properties are consistent with a role of the HPB12 protein in the structure of the B. subtilis chromosome. 相似文献
99.
The immobilization of penicillin G acylase on chitosan 总被引:3,自引:0,他引:3
100.
Permeability of human granulocytes to dimethyl sulfoxide 总被引:1,自引:0,他引:1
The permeability of the membrane of human granulocytes to the permeating solute dimethyl sulfoxide (DMSO) was studied using the Onsager form of the phenomenological equations derived from the theory of irreversible thermodynamics. Changes in cellular volume were monitored with an electronic particle counter as samples of that population were introduced into hypertonic osmotica. Temperature and concentration sensitivity analyses of the permeability coefficients were carried out. It is shown that the introduction of the Onsager formalism allows further insight into the observed transport phenomena. It was found that DMSO may affect the water permeability properties of the membrane for that population of cells. 相似文献