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101.
Kiyoshi Akeo Yasuhiko Tanaka Tatsuji Fujiwara 《In vitro cellular & developmental biology. Plant》1988,24(7):705-710
Summary The endocytotic process in cultured human RPE cells was observed after 1 min, 20 min, and 2 h incubation with cationized ferritin.
Within 1 min the ferritin particles were seen to attach to the cell membrane, especially between microvilli. Uncoated and
coated pits could be recognized on the cell membranes, and uncoated and coated endocytotic vesicles were found in the cytoplasm
after 20 min of incubation. These vesicles were surrounded by abundant microfilaments and had no visible membranes. Loss of
membrane may be an initial step in the process of developing into the irregular clumps of ferritin particles found inside
the plasma membrane. With time, more irregular clumps of ferritin, smaller than the particles introduced during incubation,
appeared just beneath the cell membrane. Lysosomes were adjacent to the clumps of ferritin particles associated with microtobules
and finally degraded these particles. The phagolysosomes containing many particles were surrounded by many microtubules. Small
ferritin particles surrounded but had not entered the rough endoplasmic reticulums, and no particles were seen either around
or in the Golgi apparatus.
Presented at the 7th International Congress of Eye Research, Nagoya, Japan, 27 September 1986. 相似文献
102.
β-lactam antibiotics in the presence of certain metal ions damage deoxyribose and DNA with the release of thiobarbituric acid-reactive material. This damage can be substantially prevented by catalase, metal chelators and some scavengers of the hydroxyl radical. Ferric salts in the presence of certain β-lactam antibiotics were effective in degrading deoxyribose but they did not appear to damage DNA. In contrast copper salts and p-lactam antibiotics were extremely effective in damaging both DNA and deoxyribose. 相似文献
103.
Mycelium ofBeauveria bassiana can be grown in liquid culture, filtered, and the mycelium dried. After rehydration the mycelium sporulates.
Two carbohydrate sources (sucrose and maltose), and one nitrogen/vitamin source (yeast extract) were tested for mycelium growth
and subsequent conidial production. Maximum mycelium growth (12.31 mg/ml), in liquid culture, was in the sucrose (3.5%)/yeast
extract (3.5%) medium, but mycelium from a maltose (2%)/yeast extract (0.75%) medium produced the maximum of 4.62×106 conidia/mg dry mycelium after incubation in moist Petri dishes.
Using the data on mycelium yield (in liquid culture) and conidial production (by dry mycelium) it is calculated that the sucrose
(3.5%)/yeast extract (3.5%) and the maltose (2%)/yeast extract (0.75%) media produce most conidia per media volume (an equivalent
of 3.52–3.72×107 conidia/ml).
相似文献
104.
Peter Zahradka 《Molecular and cellular biochemistry》1992,110(1):65-73
Summary The role of DNA polymerases in the replication of SV40 DNA was studied using a T-antigen-dependent assay supplemented with
a human KB cell extract. Inhibition of DNA polymerase α by addition of aphidicolin or monoclonal antibodies prevented DNA
synthesis, confirming the requirement for this enzyme in replication. The replication process was unaffected by ddTTP at a
concentration (5 μM) inhibitory to DNA polymerases β and γ, however, higher concentrations of ddTTP (200 μM) caused an apparent
accumulation of relaxed circular plasmid with a concomitant decrease in DNA synthesis. An analysis of this replication intermediate
indicated that it was formed during the replication reaction and that the replicative cycle was nearly complete. A kinetic
study of ddTTP inhibition strongly suggested DNA polymerase ε (PCNA-independent DNA polymerase δ) was the target of the inhibitor
and that this enzyme functions during the final stages of DNA replication. 相似文献
105.
Alessandro Negro Irene Martini Emilio Bigon Flavia Cazzola Cristina Minozzi Stephen D. Skaper Lanfranco Callegaro 《Gene》1992,110(2)
The gene(NGFB) encoding the β subunit of mature human nerve growth factor (hNGFB) was subcloned into the pJLA503 expression vector under the control of bacteriophage promoters pR and pL, and expressed in Escherichia coli. The recombinant protein represented approximately 3% of the total cellular protein. Biologically active hNGFB was solubilized (0.2% total NGFB) and purified by cation-exchange chromatography and it yielded two bands on polyacrylamide-gel electrophoresis under nonreducing conditions, corresponding to the monomeric (14 kDa) and homodimeric (26.5 kDa) forms of the molecule. Both hNGFB forms were immunopositive on Western blots with rabbit anti-NGFB antibodies; however, following additional purification, only the species corresponding to the hNGFB homodimer was biologically active on cultured chicken dorsal root ganglion neurons. These results demonstrate the feasibility of synthesizing the biologically active form of hNGFB in E. coli. 相似文献
106.
107.
In this study three different techniques suitable to recover high molecular weight genomic DNA from small blood samples of
different species of malagasy primates are compared: we suggest the use of a very simple phenol-chlorophorm DNA extraction
for badly stored or coagulated blood as for samples collected under difficult field conditions.
Furthermore we briefly describe the use of this DNA in determining RLFP patterns and DNA fingerprints. 相似文献
108.
Lubomir M. Stoilov Valeria Mirkova Jordanka Zlatanova Lalio Djondjurov 《Plant cell reports》1992,11(7):355-358
In order to elucidate some features of the topological organization of DNA within the plant nucleus, DNA fragments involved in the attachment of the DNA loops to the nuclear matrix in maize were studied. The matrix-associated DNA from dry embryo and meristematic cells after extensive digestion with DNase I and high salt treatment was about 2% of the total DNA, sized within the range of 50 and 250 bp. This DNA was found to be enriched in repetitive DNA sequences, both for nuclei from dry embryo and meristematic cells. The loop size of the DNA in cells of Zea mays appeared to be between 5 and 25 kbp.Abbreviations EDTA
Diamino-ethanetetraacetic acid
- EtBr
Ethidium bromide
- LIS
Lithium diiodosalicylate
- PMSF
Phenylmethylsulfonyl fluoride
- SDS
Sodium dodecyl sulfate 相似文献
109.
We report on the discovery and isolation of DNA- and RNA-containing macromolecular nuclear complexes whose purified major DNA possessed electrophoretic mobilities of approximately 90 and approximately 25 kbp. The deoxyribonucleoprotein-ribonucleoprotein complexes contain RNA and DNA polymerase and primase activities and were isolated from nuclei of murine RAW117 large-cell lymphoma cells by restriction digestion with Msp-I, gentle extraction with solutions containing MgCl2, but without chelating agents, and low ionic strength gel electrophoresis. Two-dimensional (isoelectric focusing/M(r)) gel electrophoresis and silver staining of the proteins of the complexes after treatment with DNase I indicated the presence of approximately 30 protein components. In vitro DNA and RNA polymerase/primase assays showed that the DNP/RNP complexes had very high enzyme specific activities. Using the DNP/RNP complexes a discrete DNA polymerase alpha product of approximately 85 kbp was synthesized that was not synthesized in the presence of the DNA polymerase alpha inhibitor aphidicolin. RNA polymerase assays in the presence of excess alpha-amanitin indicated that the complexes possessed significant RNA polymerase I activity. Preparing the complexes at various times after the release of cells from a double thymidine block showed the complexes as well as the complex-associated enzyme activities to be cell-cycle dependent. The DNA and RNA polymerase-related activities were highest in late S phase, 7 and 9 h, respectively, after release from the double thymidine block. The complexes synthesized a specific in vitro DNA polymerase product using endogenous substrate and nucleotide precursors.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
110.
Complete nucleotide sequence of the mitochondrial DNA from a liverwort,Marchantia polymorpha 总被引:1,自引:1,他引:0
Kenji Oda Katsuyuki Yamato Eiji Ohta Yasukazu Nakamura Miho Takemura Naoko Nozato Kinya Akashi Takeshi Kanegae Yutaka Ogura Takayuki Kohchi Kanji Ohyama 《Plant Molecular Biology Reporter》1992,10(2):105-163
Libraries of cosmid and plasmid clones covering the entire region of mtDNA from the liverwortMarchantia polymorpha were constructed. These clones were used for the determination of the complete nucleotide sequence of the liverwort mtDNA
totally 186,608 bp (GenBank no. M68929) and including genes for 3 species of ribosomal RNAs, 29 genes for 27 species of transfer
RNAs, and 30 genes for functionally known proteins (16 ribosomal proteins, 3 subunits of cytochromec oxidase, apocytochromeb protein, 3 subunits of H+-ATPase, and 7 subunits of NADH ubiquinone oxidoreductase). The genome also contains 32 unidentified open reading frames.
Thus the complete nucleotide sequences from both chloroplast and mitochondrial genomes have been determined in the same organism.
Plasmid clones are available upon the request.
Gene names are represented according to Lonsdale and Leaver (1988) with modifications recommended by Lonsdale (personal communication). 相似文献