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排序方式: 共有356条查询结果,搜索用时 15 毫秒
61.
62.
applications of electroporation of adherent cellsIn Situ,on a partly conductive slide 总被引:5,自引:0,他引:5
Raptis Leda H. Brownell Heather L. Liu Stanley K. W. Firth Kevin L. MacKenzie Leslie W. Stiles Charles D. Alberta John A. 《Molecular biotechnology》1995,3(2):129-134
One of the most important factors affecting the quality of PCR is the choice of primers. In general, the longer the PCR product
the more difficult it is to select efficient primers and set appropriate designing primers, and in general, the more DNA sequence
information is available, the better the ch0ance of finding an optimal primer pair. Efficient primers can be designed by avoiding
the following flaws: primer-dimer formation, self-complementarity, too lowT
m
of the primers, and/or their incorrect internal stability profile. Tips on subcloning PCR products, calculating duplex stability
(predicting dimer formation strength), and designing degenerate primers are given. 相似文献
63.
The microvessels of the pia mater lack an investment with astrocyte processes but nonetheless have a high transendothelial electrical resistance which has caused them to be regarded as part of the blood-brain barrier. This high resistance is known to be acquired in the perinatal period. The aim of our study was to relate the known physiological changes with differentiation of the endothelial paracellular clefts and especially of their tight junctions which provide the basis for the high transendothelial resistance of blood-brain barrier vessels. Tight junctions of endothelial cell paracellular clefts in pial microvessels were examined by transmission electron microscopy using goniometric tilting to reveal and measure membrane separations at tight junctions in fetal, postnatal and adult rats. These tight junctional membrane separations narrowed over the period (E16:6.3 nm, D1:6.4 nm, D7:5.4 nm) and differentiated into two groups by the adult stage: one with a membrane separation of 2.8 nm and the staining characteristics of non-brain endothelial junctions, and the other with no detectable membrane separation and the staining characteristic of blood-brain barrier endothelial junctions. This patchy and incomplete differentiation of pial tight junctions into a blood-brain barrier-like form could result either from non-uniform exposure to inductive signals or to local variation in responsiveness to such agents. Although these changes in junction organization may be related to the known increase in pial transendothelial resistance in the perinatal period, we have not yet identified any sharply defined structural change which coincides with this physiological event. 相似文献
64.
Thomas GH; Newbern EC; Korte CC; Bales MA; Muse SV; Clark AG; Kiehart DP 《Molecular biology and evolution》1997,14(12):1285-1295
Many structural, signaling, and adhesion molecules contain tandemly
repeated amino acid motifs. The alpha-actinin/spectrin/dystrophin
superfamily of F-actin-crosslinking proteins contains an array of triple
alpha-helical motifs (spectrin repeats). We present here the complete
sequence of the novel beta-spectrin isoform beta(Heavy)- spectrin (beta H).
The sequence of beta H supports the origin of alpha- and beta-spectrins
from a common ancestor, and we present a novel model for the origin of the
spectrins from a homodimeric actin-crosslinking precursor. The pattern of
similarity between the spectrin repeat units indicates that they have
evolved by a series of nested, nonuniform duplications. Furthermore, the
spectrins and dystrophins clearly have common ancestry, yet the repeat unit
is of a different length in each family. Together, these observations
suggest a dynamic period of increase in repeat number accompanied by
homogenization within each array by concerted evolution. However, today,
there is greater similarity of homologous repeats between species than
there is across repeats within species, suggesting that concerted evolution
ceased some time before the arthropod/vertebrate split. We propose a
two-phase model for the evolution of the spectrin repeat arrays in which an
initial phase of concerted evolution is subsequently retarded as each new
protein becomes constrained to a specific length and the repeats diverge at
the DNA level. This evolutionary model has general applicability to the
origins of the many other proteins that have tandemly repeated motifs.
相似文献
65.
Higher‐order assemblies of oligomeric cargo receptor complexes form the membrane scaffold of the Cvt vesicle
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Arjen J Jakobi Abul K Tarafder Yury S Bykov Andrea Picco Wanda Kukulski Jan Kosinski Wim JH Hagen Arvind C Ravichandran Matthias Wilmanns Marko Kaksonen John AG Briggs Carsten Sachse 《EMBO reports》2016,17(7):1044-1060
Selective autophagy is the mechanism by which large cargos are specifically sequestered for degradation. The structural details of cargo and receptor assembly giving rise to autophagic vesicles remain to be elucidated. We utilize the yeast cytoplasm‐to‐vacuole targeting (Cvt) pathway, a prototype of selective autophagy, together with a multi‐scale analysis approach to study the molecular structure of Cvt vesicles. We report the oligomeric nature of the major Cvt cargo Ape1 with a combined 2.8 Å X‐ray and negative stain EM structure, as well as the secondary cargo Ams1 with a 6.3 Å cryo‐EM structure. We show that the major dodecameric cargo prApe1 exhibits a tendency to form higher‐order chain structures that are broken upon interaction with the receptor Atg19 in vitro. The stoichiometry of these cargo–receptor complexes is key to maintaining the size of the Cvt aggregate in vivo. Using correlative light and electron microscopy, we further visualize key stages of Cvt vesicle biogenesis. Our findings suggest that Atg19 interaction limits Ape1 aggregate size while serving as a vehicle for vacuolar delivery of tetrameric Ams1. 相似文献
66.
In the course of a freeze-cleave study on intercellular junctions in the regenerating rat liver, we observed an unusual array of intramembranous particles located in regions of contact between endothelial cells lining the hepatic sinusoids. These arrays were characterized by an accumulation of particles which resembled a zonula occludens in their linear deployment but differed in that the contact regions were composed of individual particles which remained separated from each other by regular particle-free intervals. 相似文献
67.
The intercellular junctions of guinea-pig placental capillaries: a possible structural basis for endothelial solute permeability 总被引:1,自引:0,他引:1
The endothelial cell junction in guinea-pig placental capillaries consists of a continuous ribbon desmosome (zonula adherens) within which lies a particulate tight junction consisting of between one and five anastomosing strands. The intercellular space at these tight junctions is narrowed and is subdivided by junctional bars which are probably continuous with the intramembrane particle rows seen in freeze-fracture replicas of the junctions. Perfusion with lanthanum salts shows the gaps between the junctional bars to be lanthanum-filled and the entire junction to be lanthanum permeable. The estimated size of the spaces between the junctional bars is consistent with the junctional pore size indicated by previous ultrastructural tracer studies. The wider lateral intercellular space of the ribbon desmosome is spanned by more widely spaced "linkers" which may act as a coarser three-dimensional filter in series with size-limiting pores between the tight junctional bars. 相似文献
68.
On the Gaseous Exchange of Ammonia between Leaves and the Environment: Determination of the Ammonia Compensation Point 总被引:27,自引:6,他引:21
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Whole shoots of Phaseolus vulgaris L. and other species were exposed to a range of partial pressures of gaseous ammonia in air and the resulting fluxes were measured. Net uptake is linear with partial pressure in the range 5 to 50 nanobars and is zero at a finite partial pressure, termed the ammonia compensation point. Below the compensation point, ammonia (or possibly other volatile amines) is evolved by the leaves. The compensation points in several species are near the low partial pressures found in unpolluted air and approximate to the Km of glutamine synthetase in vitro. In P. vulgaris L., the compensation point increases with temperature. 相似文献
69.
70.
David R. Firth 《Biophysical journal》1966,6(2):201-215
Trains of nerve impulses from the crayfish stretch receptor under steady conditions are found to be extremely regular with a standard deviation of S = 0.7 x 10(-2)tau when tau, the mean interval, is in the 15 to 80 msec range. Such a small fluctuation increases the problems of film recording, accurate measurement of intervals, and especially, drifts. Experimental and mathematical techniques are described to obviate these problems. Evidence is found for a serial correlation coefficient of about -0.2 in the range, tau = 35 msec to tau = 70 msec. Shot noise and Johnson noise in a long axon are evaluated in detail and are shown to be comparable in size. It is also shown that neither shot noise nor Johnson noise is large enough to explain simply the observed interval fluctuations. Other types of membrane noise are discussed. 相似文献