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Karlheinz Grillitsch Pablo Tarazona Lisa Klug Tamara Wriessnegger Günther Zellnig Erich Leitner Ivo Feussner Günther Daum 《生物化学与生物物理学报:生物膜》2014
Despite similarities of cellular membranes in all eukaryotes, every compartment displays characteristic and often unique features which are important for the functions of the specific organelles. In the present study, we biochemically characterized the plasma membrane of the methylotrophic yeast Pichia pastoris with emphasis on the lipids which form the matrix of this compartment. Prerequisite for this effort was the design of a standardized and reliable isolation protocol of the plasma membrane at high purity. Analysis of isolated plasma membrane samples from P. pastoris revealed an increase of phosphatidylserine and a decrease of phosphatidylcholine compared to bulk membranes. The amount of saturated fatty acids in the plasma membrane was higher than in total cell extracts. Ergosterol, the final product of the yeast sterol biosynthetic pathway, was found to be enriched in plasma membrane fractions, although markedly lower than in Saccharomyces cerevisiae. A further characteristic feature of the plasma membrane from P. pastoris was the enrichment of inositol phosphorylceramides over neutral sphingolipids, which accumulated in internal membranes. The detailed analysis of the P. pastoris plasma membrane is discussed in the light of cell biological features of this microorganism especially as a microbial cell factory for heterologous protein production. 相似文献
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We have determined the canine and feline N-, K-, and H-ras gene sequences from position +23 to +270 covering exons I and II which contain the mutational hot spot codons 12, 13, and
61. The results were used to assess the degree of similarity between ras gene DNA regions containing the critical domains affected in neoplastic disorders in different mammalian species. The comparative
analyses performed included human, canine, feline, murine, rattine, and, whenever possible, bovine, leporine (rabbit), porcelline
(guinea pig), and mesocricetine (hamster) ras gene sequences within the region of interest. Comparison of feline and canine nucleotide sequences with the corresponding
regions in human DNA revealed a sequence similarity greater than 85% to the human sequence. Contemporaneous analysis of previously
published ras DNA sequences from other mammalian species showed a similar degree of homology to human DNA. Most nucleotide differences
observed represented synonymous changes without effect on the amino acid sequence of the respective proteins. For assessment
of the phylogenetic evolution of ras gene family, a maximum parsimony dendrogram based on multiple sequence alignment of the common region of exons I and II in
the N-, K-, and H-ras genes was constructed. Interestingly, a higher substitution rate among the H-ras genes became apparent, indicating accelerated sequence evolution within this particular clade. The most parsimonious tree
clearly shows that the duplications giving rise to the three ras genes must have occurred before the mammalian radiation.
Received: 23 July 1997 / Accepted: 30 October 1997 相似文献
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Anton Pauw Sunshine A. Van Bael Halton A. Peters Steven D. Allison José L. C. Camargo Miguel Cifuentes-Jara Aurlstela Conserva Teresa Garcia Restom Tamara Heartsill-Scalley Scott A. Mangan Gabriela Nunez-lturri Elsie Rivera-Ocasio Mark Rountree Susanne Vetter Carolina Volkmer de Castllho 《Biotropica》2004,36(3):410-413
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Prior intracellular recording and labeling experiments have documented local-circuit and projection neurons in the spinal trigeminal (V) nucleus with axons that arborize in more rostral and caudal spinal trigeminal subnuclei and nucleus principalis. Anterograde tracing studies were therefore carried out to assess the origin, extent, distribution, and morphology of such intersubnuclear axons in the rat trigeminal brainstem nuclear complex (TBNC). Phaseolus vulgaris leucoagglutinin (PHA-L) was used as the anterograde marker because of its high sensitivity and the morphological detail provided. Injections restricted to TBNC subnucleus caudalis resulted in dense terminal labeling in each of the more rostral ipsilateral subnuclei. Subnucleus interpolaris projected ipsilaterally and heavily to magnocellular portions of subnucleus caudalis, as well as subnucleus oralis and nucleus principalis. Nucleus principalis, on the other hand, had only a sparse projection to each of the caudal ipsilateral subnuclei. Intersubnuclear axons most frequently traveled in the deep bundles within the TBNC, the V spinal tract, and the reticular formation. They gave rise to a number of circumscribed, highly branched arbors with many boutons of the terminal and en passant types. Retrograde single- or multiple-labeling experiments assessed the cells giving rise to TBNC intersubnuclear collaterals. Horseradish peroxidase (HRP) and/or fluorescent tracer injections into the thalamus, colliculus, cerebellum, nucleus principalis, and/or subnucleus caudalis revealed large numbers of neurons in subnuclei caudalis, interpolaris, and oralis projecting to the region of nucleus principalis. Cells projecting to more caudal spinal trigeminal regions were most numerous in subnuclei interpolaris and oralis. Some cells in lamina V of subnucleus caudalis and in subnuclei interpolaris and oralis projected to thalamus and/or colliculus, as well as other TBNC subnuclei. Such collateral projections were rare in nucleus principalis and more superficial laminae of subnucleus caudalis. TBNC cells labeled by cerebellar injections were not double-labeled by tracer injections into the thalamus, colliculus, or TBNC. These findings lend generality to currently available data obtained with intracellular recording and HRP labeling methods, and suggest that most intersubnuclear axons originate in TBNC local-circuit neurons, though some originate in cells that project to midbrain and/or diencephalon. 相似文献
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Alexander I. Roitbak Elijah V. Ocherashvili Peter R. Laming Tamara A. Roitbak 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1992,170(3):327-333
Summary In 17 frogs (Rana esculenta var ridibunda) immobilised with succinyl choline the optic tectal surface was stimulated by trains of electrical pulses or by a flash to the contralateral eye. Sustained potential shifts (SPSs) and changes in extracellular potassium concentration ( [K+]0) were simultaneously recorded.In response to electrical stimulation SPSs of maximal amplitudes (1.19±0.1 mV) were recorded between 50 and 200 m in depth and maximal [K+]0 (0.69 ±0.08 mM) between 100 and 350 m. The changes of SPS and [K+]0 showed a close similarity in experiments with changes in voltage, pulse duration and frequency of stimuli within a train. The induced SPS had a duration of 28±1.54 s, the [K+]0 of 32±1.23 s.The flash stimulus induced an SPS and [K+]0 of maximal amplitudes between 50 and 200 m in depth with values of 0.57±0.1 mV and 0.29±0.03 mM respectively. An additional wave with a latency of ca 1 s and a duration of ca 3 s arose on the background of the SPS to a flash stimulus, associated with an additional increase in [K+]0.It is considered that the accumulation of K+ in extra-cellular space, with neuronal activity, results in depolarization of radial processes of ependymal glia. This is reflected in the neuropil of the upper layers of the optic tectum as an SPS.We would like to dedicate this article to the memory of Alexander Roitbak who died as a result of a tragic accident while this paper was in press. He will be remembered fondly especially for his contributions to understanding of the functions of Neuroglia. E.V.O., P.R.L., T.A.R. 相似文献
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Michel A. Haring Steve Scofield Marianne J. Teeuwen-de Vroomen Gerjan S. Leuring H. John J. Nijkamp Jacques Hille 《Plant molecular biology》1991,17(5):995-1004
A Tam3 two-element system has been designed by combining an immobilized Tam3 element with a non-autonomous dTam3 element inserted into the HPT gene. The phenotypic assay employed, restored hygromycin resistance, indicated thattrans-activation of the non-autonomous dTam3 element occurred. Molecular analyses of the excision sites revealed that the ends of the dTam3 element remain in the empty donor sites. The predominant consequence of this type of excision appears to be that excised fragments fail to re-integrate into the tobacco genome. Only one case of dTam3 re-integration could be detected. The ends of this element had been degraded upon integration into the tobacco genome. Either the altered structure of the Tam3 derivatives or tobacco host factors are influencing thetrans-activation of a dTam3 element, resulting in aberrant excision. 相似文献