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Biochemical and genetic characterization of three hamster cell mutants resistant to diphtheria toxin 总被引:2,自引:0,他引:2
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RK Draper D Chin D Eurey-Owens IE Scheffler MI Simon 《The Journal of cell biology》1979,83(1):116-125
We describe here three different hamster cell mutants which are resistant to diphtheria toxin and which provide models for investigating some of the functions required by the toxin inactivates elongation factor 2 (EF-2). Cell-free extracts from mutants Dtx(r)-3 was codominant. The evidence suggests that the codominant phenotype is the result of a mutation in a gene coding for EF-2. The recessive phenotype might arise by alteration of an enzyme which modifies the structure of EF-2 so that it becomes a substrate for reaction with the toxin. Another mutant, Dtx(r)-2, contained EF-2 that was sensitive to the toxin and this phenotype was recessive. Pseudomonas aeruginosa exotoxin is known to inactivate EF-2 as does diphtheria toxin and we tested the mutants for cross-resistance to pseudomonas exotoxin. Dtx(r)-1 and Dtx(r)-3 were cross-resistant while Dtx(r)-2 was not. It is known that diphtheria toxin does not penetrate to the cytoplasm of mouse cells and that these cell have a naturally occurring phenotype of diphtheria toxin resistance. We fused each of the mutants with mouse 3T3 cells and measured the resistance. We fused each of the mutants with mouse 3T3 cells and measured the resistance of the hybrid cells to diphtheria toxin. Intraspecies hybrids containing the genome of mutants Dtx(r)-1 and Dtx(r)-3 had some resistance while those formed with Dtx(r)-2 were as sensitive as hybrids derived from fusions between wild-type hamster cells and mouse 3T3 cells. 相似文献
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本文利用激光散斑屈光测试系统详细探讨了夜近视(night myopia)的产生机制。夜近视在环境亮度降低到产生暗视觉时出现,并随亮度继续降低而增大,平均值为1.35D。实验证明眼睛球差和色差不是造成夜近视的主要因素。夜近视数值在暗适应过程中呈增大的趋势。在暗视觉时,由于视野中缺少适宜的刺激,眼睛处于暗焦(dark focus)休止状态,这是造成夜近视的主要原因。 相似文献
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Dergacheva OIu Khachikova IE Burikov AA 《Rossi?skii fiziologicheski? zhurnal imeni I.M. Sechenova / Rossi?skaia akademiia nauk》2002,88(12):1530-1537
The nucleus pontis oralis contains several populations of neurons showing distinct sleep-waking discharge patterns. PS-on, PS-off cells, and neurons that discharged in association with phasic movements during paradoxical sleep and/or waking, were found. The findings suggest that different populations of the nucleus pontis oralis neurons take a distinct part in paradoxical sleep control. 相似文献
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本文用脑室灌注和Fura2测定细胞内游离钙技术观察了地塞米松(dexamethasone,DEX)对家兔乙二醇双(2氨基乙醚)四乙酸性发热效应和下丘脑细胞内游离钙浓度([Ca2+]i)的影响,借此深入探讨地塞米松解热作用的中枢机制。结果发现:脑室灌注乙二醇双(2氨基乙醚)四乙酸(06nmol)引起家兔结肠温度明显升高,静脉注射地塞米松(5mg/kg)显著抑制家兔乙二醇双(2氨基乙醚)四乙酸性发热,地塞米松(60~120μmol/L)并不影响下丘脑细胞内[Ca2+]i,而事先脑室灌注抑制基因转录的放线菌素D(3nmol)则完全取消了地塞米松对乙二醇双(2氨基乙醚)四乙酸性发热的解热作用。这些结果提示:地塞米松显著抑制家兔乙二醇双(2氨基乙醚)四乙酸性发热,其机制与地塞米松激活脑内某些基因的表达有关,而与下丘脑神经细胞跨膜钙离子流无关。 相似文献
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IE Hughes 《BioEssays : news and reviews in molecular, cellular and developmental biology》1999,21(11):980-981
Instant Pharmacology (1999). Saeb-Parsy K, Assomull RG, Kahn FZ, Saeb-Parsy K, and Kelly E. John Wiley and Sons 349 pp. £19.99 pbk; ISBN 0471976393 © 1999 John Wiley & Sons, Inc. 相似文献
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The distribution of the Golgi enzyme beta1, 6-N-
acetylglucosaminyltransferase (core 2 GlcNAc-T for short) has been
investigated in several tissue and cell systems by combining the potentials
of a polyclonal antibody and a novel, sensitive fluorescent enzyme assay.
In normal rat tissues, levels of the protein were found to vary and as a
general trend did not correlate with enzyme activities. Additionally, we
observed tissue-specific core 2 GlcNAc-T forms of various size: 75 kDa
(liver), 70 kDa (spleen), 60 kDA (heart), and 50 kDa (heart and lung).
These forms might arise from differential protein modifications;
alternatively, the smaller form may be a product of proteolytic cleavage,
given the presence of a catalytically inactive 50 kDa species in rat serum.
Chinese hamster ovary (CHO), MDAY-D2, PSA- 5E, and PYS-2 cell lines
consistently displayed a 70 kDa enzyme. When induced to retrodifferentiate
in the presence of butyrate + cholera toxin, CHO cells exhibited a 21-fold
increase in enzyme activity, while protein levels remained constant. A
similar trend was observed in the embryonal endoderm cell lines PSA-5E and
PYS-2, where an approximately 100-fold difference in core 2 GlcNAc-T
activity was found notwithstanding unchanged amounts of the protein and
identical mRNA levels, as evidenced by RT-PCR. In contrast, levels of core
2 GlcNAc-T activity in MDAY-D2 cells correlated well with protein
expression. Taken together, these observations demonstrate that core 2
GlcNAc-T expression may be subjected to multiple mechanisms of regulation
and suggest that in at least some instances (i.e., PSA-5E and PYS-2 cells)
expression may be regulated exclusively via posttranslational mechanism(s)
of control.
相似文献
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用 m RNA差异显示 PCR技术 ,从人 1 8周、2 2周胎儿脑和肝肾组织的 m RNA逆转录产物得到一些特异性显示的片段 .其中一个随机片段 GC1 0 2作为探针 ,从本实验室构建的 1 8周胎儿脑c DNA基因文库进行杂交筛选 ,得到一个阳性克隆λ gt1 0 /GC1 0 2 .该克隆内插入的 c DNA片段长2 .9kb,经过 DNA测序 ,显示具有一个开放阅读框架 ,编码 1 37个氨基酸的肽链 .用蛋白质结构的建模软件预测了该肽链的立体结构初步模型 . 相似文献
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基于PBL模式的生物化学教学探索 总被引:1,自引:1,他引:0
目的:评价PBL教学模式在生物化学中应用的教学效果,对生物化学理论教学的方式进行探讨。方法:用PBL模式进行教学,采用理论考试的形式对教学效果进行评价。结果:PBL教学组的学生在学习态度和积极性等方面好于传统教学组;在考试成绩方面PBL教学组病例分析考试成绩好于传统教学组,而基础知识考试成绩没有差别。结论:PBL模式能提高学生的分析问题和解决问题的综合应用能力,而基础知识的掌握程度与传统教学相比没有区别。 相似文献
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