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991.
992.
Expression of Bcl-2 protein in the epiphyseal plate cartilage and trabecular bone of growing rats 总被引:2,自引:0,他引:2
Ying Wang Renée Toury Michelle Hauchecorne N. Balmain 《Histochemistry and cell biology》1997,108(1):45-55
The protooncogene protein, Bcl-2, protects cells from apoptosis and ensures their survival in vitro by inhibiting the action
of the apoptosis-inducer, Bax. Its expression in proliferative and long-lived cells in vivo also indicates that it protects
against cell death. The chondrocytes of the epiphyseal plate cartilage undergo a series of maturation steps and deposit mineral
in the cartilage matrix before dying. The possibility that Bcl-2 helps protect chondrocytes until mineral deposition is completed
was investigated by determining the distribution of Bcl-2 immunoreactivity in the epiphyseal plate cartilage of growing rats
and its subcellular localization, using a specific antibody. The involvement of Bax in the triggering of chondrocyte death
was checked by immunocytochemistry. Bcl-2 expression in the osteoblasts and the final result of their evolution, the osteocytes,
was also examined in trabecular bone. Bcl-2 immunoreactivity was non-uniformly distributed throughout the epiphyseal cartilage.
It was maximal in proliferative chondrocytes, decreased in mature chondrocytes, and low in hypertrophic chondrocytes, whereas
there was Bax immunoreactivity in all chondrocytes examined. Immunolabeling was intense in osteoblasts but considerably lower
in fully differentiated osteocytes. Bcl-2 immunoreactivity was mainly in the cytoplasm of chondrocytes, osteoblasts, and early
osteocytes; the nuclei appeared clear. The subcellular distribution of Bcl-2 immunolabeling in chondrocytes, revealed by gold
particles in the electron microscope, showed that gold particles were frequently concentrated in the mitochondria in all the
cartilage zones and lay mainly within the organelles, not at their periphery. The endoplasmic reticulum contained moderate
immunoreactivity and there were few gold particles in the cytoplasm and nuclei. The number of gold particles decreased in
all the subcellular compartments from proliferative to hypertrophic chondrocytes. In contrast, Bax immunoreactivity changed
little during chondrocyte terminal evolution, and its subcellular distribution mirrored that of Bcl-2. These immunocytochemical
data indicate that Bcl-2 helps maintain chondrocytes and osteoblasts until their terminal maturation.
Accepted: 19 February 1997 相似文献
993.
994.
995.
Silica gel TLC methods were developed for the separation of 2,4,6-trinitrotoluene (TNT) in mixtures with possible reduction products. The methods employed repeated elutions with simple binary or ternary solvent systems in either one or two dimensional modes. The resolved analytes include TNT, selected amino derivatives (2-amino-4,6-di-nitrotoluene, 4-amino-2,6-dinitrotoluene, 2,4-diamino-6-nitrotoluene) and known hydroxylamino derivatives (2-hydroxyl-amino-4,6-dinitrotoluene, 4-hydroxylamino-2,6-dinitrotoluene and 2,4-dihydroxylamino-6-nitrotoluene). 相似文献
996.
Michel Dion Daniel Charlier Haifeng Wang Daniel Gigot Alexey Savchenko Jean-Noël Hallet Nicolas Glansdorff & Vehary Sakanyan 《Molecular microbiology》1997,25(2):385-398
We report here the cloning of the arginine repressor gene argR of Bacillus stearothermophilus and the characterization and purification to homogeneity of its product. The deduced amino acid sequence of the 16.8-kDa ArgR subunit shares 72% identity with its mesophilic homologue AhrC of Bacilus subtilis . Sequence analysis of B. stearothermophilus ArgR and comparisons with mesophilic arginine repressors suggest that the thermostable repressor comprises an N-terminal DNA-binding and a C-terminal oligomerization and arginine-binding region. B. stearothermophilus ArgR has been overexpressed in E. coli and purified as a 48.0-kDa trimeric protein. The repressor inhibits the expression of a B. stearothermophilus argC–lacZ fusion in E. coli cells. In the presence of arginine, the purified protein binds tightly and specifically to the argC operator, which largely overlaps the argC promoter. The purified B. stearothermophilus repressor proved to be very thermostable with a half-life of approximately 30 min at 90°C, whereas B. subtilis AhrC was largely inactivated at 65°C. Moreover, ArgR operator complexes were found to be remarkably thermostable and could be formed efficiently at up to 85°C, well above the optimal growth temperature of the moderate thermophile B. stearothermophilus . This pronounced resistance of the repressor–operator complexes to heat treatment suggests that the same type of regulatory mechanism could operate in extreme thermophiles. 相似文献
997.
Q. Wang C.K. Noguchi M. Kuninobu Y. Hara K. Kakimoto H.I. Ogawa Y. Kato 《Biotechnology Techniques》1997,11(2):105-108
Continuous anaerobic digestion of waste activated sludge pretreated at low temperatures below 100°C increased methane generation by 30%. pH values of the digestion mixture increased, approximately from 0.3 to 0.55 by pretreatment, although its volatile fatty acids concentration was greater than the control. An abrupt increase in propionate : acetate ratio in digestion stage (e.g. from less than 1:1 to over 3.5 :1), provided a reliable indicator for impending failure.This revised version was published online in October 2005 with corrections to the Cover Date. 相似文献
998.
Agrobacterium tumefaciens-mediated barley transformation 总被引:26,自引:2,他引:24
Sonia Tingay David McElroy Roger Kalla Sarah Fieg Mingbo Wang Sarah Thornton Richard Brettell 《The Plant journal : for cell and molecular biology》1997,11(6):1369-1376
Genetically transformed barley was produced by eco-cultivating immature embryo explants with Agrobacterium tumefaciens carrying a binary vector coding for chimaeric bacterial genes, bar and gus , and selecting for bialaphos-resistant cultures from which plants were regenerated. Integration of both genes was confirmed by gel blot hybridization analysis of DNA from the transformed plants and their progenies. From 1282 embryos, plants were recovered for 54 independently transformed lines, giving a transformation efficiency of 4.2%. Transgene numbers in the different lines ranged from single copy insertion to at least ten copies. Sixteen out of 18 plants grown to maturity were fully fertile. Both marker genes, bar and gus , were expressed and co-segregated in the T1 progeny plants. In the majority of cases, the genes showed Mendelian segregation predicted for transgene insertion at a single locus. In one family with multiple transgene insertions, molecular analysis of T1 and T2 plants suggested that the T-DNA had inserted at two unlinked loci. 相似文献
999.
Ming Li Wang Lu Huang Deverie K. Bongard-Pierce Stephen Belmonte Eve Ann Zachgo John W. Morris Maureen Dolan Howard M. Goodman 《The Plant journal : for cell and molecular biology》1997,12(3):711-730
A method for construction of bacterial artificial chromosome (BAC) contigs from a yeast artifical chromosome (YAC) physical map is described. An ∼2 Mb contig, consisting of two large BAC contigs linked by a small YAC, has been assembled in the region around 80 cM of Arabidopsis thaliana chromosome 2. Clones from this contig will facilitate gene isolation in the region and can be used directly as substrates for DNA sequencing. 相似文献
1000.
Conformation of P22 tailspike folding and aggregation intermediates probed by monoclonal antibodies. 总被引:3,自引:2,他引:1 下载免费PDF全文
M. A. Speed T. Morshead D. I. Wang J. King 《Protein science : a publication of the Protein Society》1997,6(1):99-108
The partitioning of partially folded polypeptide chains between correctly folded native states and off-pathway inclusion bodies is a critical reaction in biotechnology. Multimeric partially folded intermediates, representing early stages of the aggregation pathway for the P22 tailspike protein, have been trapped in the cold and isolated by nondenaturing polyacrylamide gel electrophoresis (PAGE) (speed MA, Wang DIC, King J. 1995. Protein Sci 4:900-908). Monoclonal antibodies against tailspike chains discriminate between folding intermediates and native states (Friguet B, Djavadi-Ohaniance L, King J, Goldberg ME. 1994. J Biol Chem 269:15945-15949). Here we describe a nondenaturing Western blot procedure to probe the conformation of productive folding intermediates and off-pathway aggregation intermediates. The aggregation intermediates displayed epitopes in common with productive folding intermediates but were not recognized by antibodies against native epitopes. The nonnative epitope on the folding and aggregation intermediates was located on the partially folded N-terminus, indicating that the N-terminus remained accessible and nonnative in the aggregated state. Antibodies against native epitopes blocked folding, but the monoclonal directed against the N-terminal epitope did not, indicating that the conformation of the N-terminus is not a key determinant of the productive folding and chain association pathway. 相似文献