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131.
M A Chan L D Stein H M Dosch N H Sigal 《Journal of immunology (Baltimore, Md. : 1950)》1986,136(1):106-112
Although most human B cells express receptors for Epstein Barr virus (EBV), few (usually less than 1%) are readily transformed into B lymphoblastoid cell lines after exposure to EBV. Transformable cells previously have been found to be mostly resting B lymphocytes. We recently developed a limiting dilution culture system which permits the growth of EBV-transformed B lymphocytes with high efficiency. Because in this system up to over 30% of peripheral blood- or tonsil-derived B cells respond to EBV, we re-examined the properties of EBV-transformable cells. Frequencies of transformable lymphocytes were determined by Poisson analysis. EBV-susceptible B cells committed to IgM, IgG, or IgA secretion were found to occur in the range of 3 to 27, 0.1 to 6, and 0.1 to 5 per 100 B cells, respectively. Under our culture conditions, a major proportion of the IgM-committed cells derived from large lymphocytes which appeared to have entered the cell cycle. This population contains most of the EBV-responsive cells detected and, therefore, most of the additional cells responding in our culture system. In contrast, precursors of IgG- or IgA-producing lymphoblast lines were small cells with DNA contents typical for the G0/G1 phases of the cell cycle. Anti-immunoglobulin antibodies were used in panning experiments to separate B cell subpopulations which expressed different immunoglobulin isotypes on their surface. In limiting dilution cultures of these purified B lymphocytes subsets, it was found that virtually all precursors of IgM-producing cell lines expressed surface IgM (sIgM) before their infection and transformation by EBV. The "cloning efficiency" of positively selected, large sIgM+ cells approached 100%. In contrast, sIgG or sIgA were found only on cells committed to the production of IgG or IgA, respectively. The expression of sIgD was examined by using sequential panning procedures. Virtually all IgM-committed lymphocytes and a subset of cells committed to IgA secretion were found among the sIgD+ cells. The majority of cells committed to IgA production and all IgG-committed cells were found in the sIgD- B cell population. Our findings indicate that the EBV-susceptible B cell subset of normal lymphocytes is heterogeneous with respect to cell size, cell cycle, sIg determinants, and efficiency of transformation. On the basis of our findings and previous reports, we propose a model in which transformability is a B cell-inherent property. Factors unrelated to the virus but present in our culture system appear responsible for the enhanced vulnerability to viral transformation in some cells which entered into the cell cycle.(ABSTRACT TRUNCATED AT 400 WORDS) 相似文献
132.
Transient response of forests to CO2-induced climate change: simulation modeling experiments in eastern North America 总被引:1,自引:0,他引:1
Allen M. Solomon 《Oecologia》1986,68(4):567-579
Summary The temporal response of forests to CO2-induced climate changes was examined for eastern North America. A forest stand simulation model was used with the assumption that climate will change at a constant rate as atmospheric CO2 doubles, and then as CO2 doubles again. Before being used to project future vegetation trends, the simulation model FORENA was verified by its ability to reproduce long, temporal sequences of plant community change recorded by fossil pollen and by its ability to reproduce today's vegetation. The simulated effects of changing monthly temperature and precipitation included a distinctive dieback of extant trees at most locations, with only partial recovery of biomass in areas of today's temperate deciduous forest. In the southern portion of today's deciduous-coniferous transition forests the simulated dieback was indistinct and recovery by deciduous tree species was rapid. In more northerly transition areas, the dieback not only was clearly expressed, but occurred twice, when new dominant species replaced extant conifers, then were themselves replaced, as climate change continued. Boreal conifers also underwent diebacks and were replaced by deciduous hardwoods more slowly in the north than in the south. Transient responses in species composition and carbon storage continued as much as 300 years after simulated climate changes ceased.Environmental Sciences Division Publication No. 2625 相似文献
133.
Caitilyn Allen Verlyn K. Stromberg Franzine D. Smith George H. Lacy Mark S. Mount 《Molecular & general genetics : MGG》1986,202(2):276-279
Summary We report the complementation of a genetic lesion in the genome of Erwinia carotovora subsp. carotovora (Ecc), a pathogenic bacterium that incites soft rot of plants. A Sau3AI genomic library of Ecc was constructed using the conjugal cosmid pLAFR-3 as a vector. Sixteen cosmid clones encoding various plant tissue-degrading enzymes were identified, including a proteolytic clone, five cellulolytic clones, and ten pectolytic clones. We detected a mutant of Ecc with no proteolytic activity following transposon mutagenesis with an unstable Tn5-carrying plasmid. Conjugal transfer of the protease-encoding cosmid to this mutant restored near-wildtype extracellular protease production. Further manipulation and study of genes encoding pathogenic determinants in Ecc will be possible using this system. 相似文献
134.
Growth, dry matter partitioning, and diurnal activities of RuBP carboxylase in citrus seedlings maintained at two levels of CO2 总被引:3,自引:0,他引:3
Karen E. Koch Pierce H. Jones Wayne T. Avigne L. Hartwell Allen Jr. 《Physiologia plantarum》1986,67(3):477-484
The long-term response of citrus rootstock seedlings to CO2 enrichment was examined in Carrizo estrange ( Poncirua trifoliata (L.) Raf. x Citrus sinensis (L.) Osbeck] and Swingle citrumelo ( P. trifoliate x C. parodist Macf.]. Plaotlets 14 weeks old were transferred to outdoor controlled-environment chambers and maintained for 5 months from Feb. 14 to July 21. During this period, new growth (cm) of citrange and citrumelo shoots at 660 μl1−1 was 94 and 69% greater, respectively, than at 330 μ1 1−1 . Total dry weight of both rootstock shoots had increased by over 100%. Growth of few species is affected this markedly by elevated CO2 levels.
More carbon was partitioned to above-ground organs in CO2 -enriched citrus seedlings. Stem dry matter per unit length was also 32 and 44% greater in citrange and citrumelo, respectively. Total leaf area was increased by 124% in citrange and 85% in citrumelo due to greater leaf number and size. Variations in overall relative growth rate appeared to be related to the rapid, sequential, flush-type growth in citrus, in which an entire shoot segment with its associated leaves remains an active sink until fully expanded. RuBP carboxylase (EC 4.1.1.39) activity in leaves of recently-expanded flushes was higher in citrumelo plants grown at 660 vs 330 μ1 1−1 CO2 and changed diurnally for citrange (but not citrumelo) leaves at both CO2 levels. The results are consistent with the hypothesis that positive long-term effects of CO2 enrichment may be greater in species or during growth periods where sink capacity for carbon utilization is high. 相似文献
More carbon was partitioned to above-ground organs in CO
135.
Cloning, nucleotide sequencing, and expression of tetanus toxin fragment C in Escherichia coli. 总被引:14,自引:0,他引:14 下载免费PDF全文
N F Fairweather V A Lyness D J Pickard G Allen R O Thomson 《Journal of bacteriology》1986,165(1):21-27
The amino acid sequence of the first 30 residues of fragment C of tetanus toxin was determined, and a mixture of 32 complementary oligonucleotides, each 17 bases long, was synthesized. A 2-kilobase (kb) EcoI fragment of Clostridium tetani DNA was identified by Southern blotting and was cloned into the Escherichia coli plasmid vector pAT153 with the 32P-labeled oligonucleotide mixture as a probe. A second 3.2-kb Bg/II fragment was identified and cloned with the 2-kb EcoRI fragment as a probe. The nucleotide sequence of 1.8 kb of this DNA was determined and was shown to encode the entire fragment C and a portion of fragment B of tetanus toxin. The tetanus DNA was expressed in E. coli with pWRL507, a plasmid vector containing the trp promoter and a portion of the trpE gene. The trpE-tetanus fusion proteins were visualized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and were shown to react with anti-fragment C antibody. 相似文献
136.
M Setaka N Satoh T Kobayashi T Hongo T Kwan A Yamaguchi M Futai 《Journal of biochemistry》1986,99(3):777-783
A phospholipid bilayer membrane was spread from an organic solvent solution between a polyacrylamide gel surface and an aqueous buffer solution. The membrane was quite similar to the conventional black lipid membrane, but was of a large size and was stable since it was supported on the gel surface. Bacteriorhodopsin, impregnated into the membrane, generated membrane potential and current upon illumination. The induced current was large, and this was attributed to the large area of the present membrane. Remarkable responses of the light-induced potential and current were also observed with a thick layer of organic solvent containing phospholipids. The effects of applied membrane potential, carbonylcyanide-m-chlorophenyl hydrazone (CCCP) and gramicidin were examined on these photoresponses. Steady-state current, which is due to protons flowing through the membrane, was enormously enhanced by applying membrane potential opposite to the photopotential or by adding gramicidin to the membrane-forming solution. 相似文献
137.
Promoter recognition by Escherichia coli RNA polymerase. Effects of substitutions in the spacer DNA separating the -10 and -35 regions 总被引:16,自引:0,他引:16
A family of variants of the PRM promoter of lambda phage was constructed, bearing nine base pair substitutions in a stretch of the spacer DNA separating the contacted -10 and -35 regions. The substituted sequences were chosen for their potential to adopt structures different from those of average B-form DNA and thus to affect the interaction of RNA polymerase with the two contacted regions. Characterization of the promoters in vitro and in vivo provides additional support for the lack of specific contacts in the substituted spacer region and shows that a small change in the relative rotational orientation of the -10 and -35 regions is inconsequential to promoter function. However, a 2-3-fold reduction in promoter activity is observed with promoters bearing substitutions of nonalternating dG-dC base pairs in either orientation. This corroborates other studies indicating the anomalous behavior of such sequences and suggests that the structure of the spacer DNA can modulate promoter recognition. 相似文献
138.
Chemical and spectroscopic evidence for the formation of a ferryl Fea3 intermediate during turnover of cytochrome c oxidase 总被引:1,自引:0,他引:1
When partially reduced cytochrome c oxidase samples are reoxidized with dioxygen, an EPR-silent dioxygen intermediate, which is at the three-electron level of dioxygen reduction, is trapped at the dioxygen reduction site. The intermediate has novel spectral features at 580 and 537 nm. Combined optical and EPR results reveal that this intermediate reacts rapidly with CO at 277-298 K causing the abolition of the 580/537 mm features and the appearance of a rhombic CuB EPR signal. A ferryl Fea3, or an intermediate at the same formal level of oxidation, is proposed to oxidize CO to CO2 producing an EPR-detectable CuB adjacent to a low-spin ferrous Fea3-dioxygen (or carbon monoxide) adduct. 相似文献
139.
Brain peptides and glial growth. I. Glia-promoting factors as regulators of gliogenesis in the developing and injured central nervous system 总被引:7,自引:2,他引:5 下载免费PDF全文
Glia-promoting factors (GPFs) are peptides of the central nervous system which accelerate the growth of specific glial populations in vitro. Although these factors were first discovered in the goldfish visual system (Giulian, D., Y. Tomozawa, H. Hindman, and R. Allen, 1985, Proc. Natl. Acad. Sci. USA., 83:4287-4290), we now report similar peptides are found in mammalian brain. The cerebral cortex of rat contains oligodendroglia-stimulating peptides, GPF1 (15 kD) and GPF3 (6 kD), as well as astroglia-stimulating peptides, GPF2 (9 kD) and GPF4 (3 kD). The concentrations of specific GPFs increase in brain during periods of gliogenesis. For example, GPF1 and GPF3 are found in postnatal rat brain during a peak of oligondendroglial growth while GPF2 and GPF4 are first detected at a time of astroglial proliferation in the embryo. Stab wound injury to the cerebral cortices of rats stimulates astroglial proliferation and induces marked elevations in levels of GPF2 and GPF4. Our findings suggest that two distinct classes of GPFs, those acting upon oligodendroglia and those acting upon astroglia, help to regulate cell growth in the developing and injured central nervous system. 相似文献
140.
Characterization of the phosphotyrosyl protein phosphatase activity of calmodulin-dependent protein phosphatase 总被引:4,自引:0,他引:4
C P Chan B Gallis D K Blumenthal C J Pallen J H Wang E G Krebs 《The Journal of biological chemistry》1986,261(21):9890-9895
Calmodulin-dependent protein phosphatase from bovine brain and heart was assayed for phosphotyrosine and phosphoserine phosphatase activity using several substrates: 1) smooth muscle myosin light chain (LC20) phosphorylated on tyrosine or serine residues, 2) angiotensin I phosphorylated on tyrosine, and 3) synthetic phosphotyrosine- or phosphoserine-containing peptides with amino acid sequences patterned after the autophosphorylation site in Type II regulatory subunit of the cAMP-dependent protein kinase. The phosphatase was activated by Ni2+ and Mn2+, and stimulated further by calmodulin. In the presence of Ni2+ and calmodulin, it exhibited similar kinetic constants for the dephosphorylation of phosphotyrosyl LC20 (Km = 0.9 microM, and Vmax = 350 nmol/min/mg) and phosphoseryl LC20 (Km = 2.6 microM, Vmax = 690 nmol/min/mg). Dephosphorylation of phosphotyrosyl LC20 was inhibited by phosphoseryl LC20 with an apparent Ki of 2 microM. Compared to the reactions with phosphotyrosyl LC20 as the substrate, reactions with phosphotyrosine-containing oligopeptides exhibited slightly higher Km and lower Vmax values. The reaction with the phosphoseryl peptide based on the Type II regulatory subunit sequence exhibited a slightly higher Km (23 microM), but a much higher Vmax (4400 nmol/min/mg) than that with its phosphotyrosine-containing counterpart. Micromolar concentrations of Zn2+ inhibited the phosphatase activity; vanadate was less potent, and 25 mM NaF was ineffective. The study provides quantitative data to serve as a basis for comparing the ability of the calmodulin-dependent protein phosphatase to act on phosphotyrosine- and phosphoserine-containing substrates. 相似文献