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1.
Richard E. Kouri Raymond Kiefer Eugene M. Zimmerman 《In vitro cellular & developmental biology. Plant》1974,10(1-2):18-25
Summary Two methods for determining the hydrocarbon-metabolizing enzyme activity of cultured mammalian cells were compared. The method
designed to measure benzo[a]an-thracene-induced aryl hydrocarbon hydroxylase activity could detect and quantify enzyme activities
in low passage rodent cells, but could not reproducibly detect levels in intermediate or high passage mouse, rat, or human
cells. The method designed to measure the ability of a cell to convert benzo[a]pyrene from an organic-soluble to an aqueous
acetone-soluble form proved to be more reproducible. This technique, when modified, was demonstrated to be an effective screening
test for the detection of those lines with higher levels of hydrocarbon-metabolizing enzymes.
Supported by the Council for Tobacco Research and Contract NIH 70-2068 within the Virus Cancer Program, National Cancer Institute,
National Institutes of Health. 相似文献
2.
F. Fontaine E. Kiefer C. Clément M. Burrus J. L. Druelle 《Trees - Structure and Function》1999,14(2):83-90
In the present work, we described the fate of proventitious epicormic buds on the trunks of 40-year-old Quercus petraea trees and in parallel the vascular trace they produced in the wood. Our results show that small and large individual epicormic
buds can survive as buds for 40 years and that both are composed of a terminal meristem and scales. Meristematic areas are
detected in the scale axils of small buds; in addition to these meristems the large buds also have secondary bud primordia.
The small buds are connected to the pith of the main stem by a unique trace, whereas the large buds are connected by one or
multiple traces. A single trace might imply that the whole bud is still alive and multiple traces might indicate that the
terminal meristem has died. In the latter case, each trace is connected to a secondary bud of the large bud. The buds found
in a cluster are composed of a terminal meristem and scales with axillary meristems in the scale axils. A cluster is connected
to the pith of a stem either by a unique trace when it seems to be the result of partial abscission of an epicormic shoot
or multiple traces when it might have originated from an epicormic bud in which the terminal meristem has died. Whatever the
type of the bud, the vascular trace in the bark is composed of a cambium, secondary xylem and parenchyma cells and the trace
present in the wood had parenchyma cells with vestiges of secondary xylem. Each year, the vascular trace should be produced
in the bark by the cambium of the tree but not by the bud itself. On 40-year-old Q. petraea, we observed a proliferation of epicormic buds and in parallel a multiplication of the number of vascular traces in the trunk,
but the knots caused by the traces of epicormic buds in the wood, either as individuals or in clusters, are minor since their
colours are only slightly darker than those of woody rays and they are less than 2 mm in diameter. The knots will appear when
epicormic buds develop into shoots.
Received: 30 March 1999 / Accepted: 09 June 1999 相似文献
3.
M P Neeper L M Kuo M C Kiefer R J Robb 《Journal of immunology (Baltimore, Md. : 1950)》1987,138(10):3532-3538
High affinity receptors for interleukin 2 (IL 2) contain the Tac protein as one ligand-binding subunit. Localization of the IL 2-binding site on this molecule, as well as localization of the complementary site on IL 2, should provide insight into the design of IL 2 analogs. In this report, we examine the ability of normal and modified Tac protein to bind IL 2 and several antibodies that recognize the native Tac molecule. Using a transient L cell expression system, we have determined that transfection with cDNA-missing Tac exon 4 resulted in expression of spliced protein that had no measurable binding to IL 2 or the monoclonal anti-receptor antibodies, anti-Tac, and 7G7/B6. This protein was detected, however, by rabbit polyclonal antibodies prepared against synthetic Tac peptides. Thus, one or more amino acids encoded by exon 4 is important, either for direct ligand contact or for the proper folding of critical segments of the Tac molecule. In addition, insertion of stop codons at a unique restriction enzyme site near the beginning of exon 5 resulted in cellular secretion of truncated Tac molecules that were capable of binding IL 2, anti-Tac, and 7G7/B6. Amino acids encoded by exons 5 to 8 thus play no critical role in IL 2 binding. The ligand association demonstrated for truncated Tac protein produced by exons 2 to 4 should guide attempts to define the IL 2-binding segment of the Tac molecule. 相似文献
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7.
P J Silver R E Lepore W R Cumiskey D Kiefer A L Harris 《Biochemical and biophysical research communications》1988,154(1):272-277
Protein kinase C (PKC) activity in aortic and renal arterial smooth muscle from SHR (20-23 wk male; mean arterial pressure = 178 mm Hg) and WKY (age/sex matched; mean arterial pressure = 126 mm Hg) was quantitated. Activity was greatest in the particulate fractions relative to the soluble fractions in all sources. The only difference between SHR and WKY was in the soluble fraction from SHR renal arteries, which had 2 fold more activity (255 pmol/mg/min) when compared with WKY (136 pmol/mg/min). This difference was not apparently related to force modulation, since the magnitude of isometric force development in renal arteries in response to phorbol 12,13-dibutyrate was not different between SHR and WKY. The magnitude of force developed in response to phorbol 12,13-dibutyrate and PKC activity in the particulate fraction was greatest in aorta vs. renal arteries in both WKY and SHR. These results suggest that regional vascular differences in the amount of PKC activity may exist which are not apparently related to a disease state (i.e., hypertension). These differences may be related to differential sensitivity to phorbol ester-mediated contractions in isolated smooth muscle. 相似文献
8.
Isolation of mouse N-CAM-related cDNA: detection and cloning using monoclonal antibodies. 总被引:28,自引:5,他引:23 下载免费PDF全文
C Goridis M Hirn M J Santoni G Gennarini H Deagostini-Bazin B R Jordan M Kiefer M Steinmetz 《The EMBO journal》1985,4(3):631-635
Clones coding for the mouse neural cell adhesion molecule (N-CAM) were isolated from a cDNA library prepared in the expression vector lambda gt 11 from mRNA extracted from a mouse neuroblastoma cell line. This library was screened with two anti-N-CAM monoclonal antibodies directed against different sites on the molecule and with rabbit anti-N-CAM serum. Two clones were identified with the first monoclonal antibody, three with the second one, none reacted with both. The relevance of these cDNA clones to N-CAM was confirmed by several observations. First, cDNA sequences detected with one monoclonal antibody cross-hybridized with those identified by the other antibody. Second, the different fusion proteins all bound the rabbit serum in addition to one monoclonal antibody. Finally, the probes hybridized to discrete mRNA species of sufficient lengths to code for the very large N-CAM polypeptides in RNA preparations from N-CAM-expressing, but not from N-CAM-negative cells. An additional mRNA species not seen in embryonic brain was expressed in adult mouse brain. Genomic blot experiments indicated that sequences corresponding to one of our probes are present only a few times in the mouse genome. 相似文献
9.
A comparative description of mitochondrial DNA differentiation in selected avian and other vertebrate genera 总被引:14,自引:1,他引:13
Levels of mitochondrial DNA (mtDNA) sequence divergence between species
within each of several avian (Anas, Aythya, Dendroica, Melospiza, and
Zonotrichia) and nonavian (Lepomis and Hyla) vertebrate genera were
compared. An analysis of digestion profiles generated by 13-18 restriction
endonucleases indicates little overlap in magnitude of mtDNA divergence for
the avian versus nonavian taxa examined. In 55 interspecific comparisons
among the avian congeners, the fraction of identical fragment lengths (F)
ranged from 0.26 to 0.96 (F = 0.46), and, given certain assumptions, these
translate into estimates of nucleotide sequence divergence (p) ranging from
0.007 to 0.088; in 46 comparisons among the fish and amphibian congeners, F
values ranged from 0.00 to 0.36 (F = 0.09), yielding estimates of P greater
than 0.070. The small mtDNA distances among avian congeners are associated
with protein-electrophoretic distances (D values) less than approximately
0.2, while the mtDNA distances among assayed fish and amphibian congeners
are associated with D values usually greater than 0.4. Since the
conservative pattern of protein differentiation previously reported for
many avian versus nonavian taxa now appears to be paralleled by a
conservative pattern of mtDNA divergence, it seems increasingly likely that
many avian species have shared more recent common ancestors than have their
nonavian taxonomic counterparts. However, estimates of avian divergence
times derived from mtDNA- and protein-calibrated clocks cannot readily be
reconciled with some published dates based on limited fossil remains. If
the earlier paleontological interpretations are valid, then protein and
mtDNA evolution must be somewhat decelerated in birds. The empirical and
conceptual issues raised by these findings are highly analogous to those in
the long-standing debate about rates of molecular evolution and times of
separation of ancestral hominids from African apes.
相似文献
10.
Methods for computing the standard errors of branching points in an evolutionary tree and their application to molecular data from humans and apes 总被引:23,自引:2,他引:21
Statistical methods for computing the standard errors of the branching
points of an evolutionary tree are developed. These methods are for the
unweighted pair-group method-determined (UPGMA) trees reconstructed from
molecular data such as amino acid sequences, nucleotide sequences,
restriction-sites data, and electrophoretic distances. They were applied to
data for the human, chimpanzee, gorilla, orangutan, and gibbon species.
Among the four different sets of data used, DNA sequences for an
895-nucleotide segment of mitochondrial DNA (Brown et al. 1982) gave the
most reliable tree, whereas electrophoretic data (Bruce and Ayala 1979)
gave the least reliable one. The DNA sequence data suggested that the
chimpanzee is the closest and that the gorilla is the next closest to the
human species. The orangutan and gibbon are more distantly related to man
than is the gorilla. This topology of the tree is in agreement with that
for the tree obtained from chromosomal studies and DNA-hybridization
experiments. However, the difference between the branching point for the
human and the chimpanzee species and that for the gorilla species and the
human-chimpanzee group is not statistically significant. In addition to
this analysis, various factors that affect the accuracy of an estimated
tree are discussed.
相似文献