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通过改良贝伦斯技术分离得到的细胞核的游离氨基酸和核酸含量

Free amino acids and nucleic acid content of cell nuclei isolated by a modification of Behrens' technique.

作者信息

DOUNCE A L, TISHKOFF G H, BARNETT S R, FREER R M

出版信息

J Gen Physiol. 1950 May 20;33(5):629-42. doi: 10.1085/jgp.33.5.629.

DOI:10.1085/jgp.33.5.629
PMID:15422114
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2147208/
Abstract
  1. Nuclei were prepared from frozen rat liver by a modification of the technique of Behrens, and were studied with regard to the content of free amino acids and nucleic acid. 2. Under rigorously controlled conditions, preparations of nuclei are obtained by the Behrens' method which form a gel in the presence of 5 or 10 per cent NaCl or of water plus a small amount of dilute alkali; whereas when conditions are less rigorously controlled, nuclei are obtained which form no such gel. The property of forming gels with alkali is probably characteristic of all cell nuclei which have not undergone autolysis. 3. Nuclei prepared by the Behrens' technique contain the enzymes arginase, catalase, and esterase in very appreciable concentrations. 4. The free amino acids of the isolated cell nuclei, as well as of other liver cell fractions, have been investigated using the technique of paper chromatography. 5. The chromatographic patterns of the free amino acids of whole cells, ground cytoplasm, and isolated cell nuclei were very similar or identical. A feature of interest in these chromatograms was the faintness or absence of the spots due to a number of the essential amino acids, as compared to the intensities of the spots due to glycine, alanine, and glutamic acid. Glutathione was present in the isolated nuclei as well as in the whole cells. 6. Chromatograms made from hydrolysates of nuclei showed high concentrations of the essential amino acids and were similar to chromatograms of hydrolysates of typical proteins.
摘要
  1. 通过对贝伦斯技术进行改进,从冷冻大鼠肝脏中制备细胞核,并对其游离氨基酸和核酸含量进行研究。2. 在严格控制的条件下,采用贝伦斯方法可获得细胞核制剂,该制剂在5%或10%氯化钠存在下,或在水加少量稀碱的情况下会形成凝胶;而当条件控制不那么严格时,获得的细胞核不会形成这种凝胶。用碱形成凝胶的特性可能是所有未发生自溶的细胞核的特征。3. 用贝伦斯技术制备的细胞核含有浓度相当可观的精氨酸酶、过氧化氢酶和酯酶。4. 利用纸层析技术对分离的细胞核以及其他肝细胞组分中的游离氨基酸进行了研究。5. 全细胞、研磨后的细胞质和分离的细胞核中游离氨基酸的层析图谱非常相似或相同。这些层析图谱中一个有趣的特征是,与甘氨酸、丙氨酸和谷氨酸产生的斑点强度相比,许多必需氨基酸产生的斑点较淡或没有。谷胱甘肽在分离的细胞核以及全细胞中均有存在。6. 由细胞核水解产物制成的层析图谱显示必需氨基酸浓度很高,且与典型蛋白质水解产物的层析图谱相似。

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Free amino acids and nucleic acid content of cell nuclei isolated by a modification of Behrens' technique.通过改良贝伦斯技术分离得到的细胞核的游离氨基酸和核酸含量
J Gen Physiol. 1950 May 20;33(5):629-42. doi: 10.1085/jgp.33.5.629.
2
Studies of two types of alkaline phosphatase in nuclei isolated from the livers of fed and fasted rats by a modification of the Behrens technique.通过对贝伦斯技术进行改进,对从喂食和禁食大鼠肝脏中分离出的细胞核中的两种碱性磷酸酶进行研究。
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A method for isolating intact mitochondria and nuclei from the same homogenate, and the influence of mitochondrial destruction on the properties of cell nuclei.一种从同一匀浆中分离完整线粒体和细胞核的方法,以及线粒体破坏对细胞核性质的影响。
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J Gen Physiol. 1953 Nov 20;37(2):177-87. doi: 10.1085/jgp.37.2.177.

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Chloroplast Isolation in Nonaqueous Media.非水介质中叶绿体的分离
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3
The Physical Environment of Chloroplasts as Related to their Morphology and Activity in Vitro.叶绿体的物理环境与其体外形态和活性的关系
Plant Physiol. 1954 Sep;29(5):448-58. doi: 10.1104/pp.29.5.448.
4
AMINO ACID ACTIVATION AND TRANSFER TO RIBONUCLEIC ACIDS IN THE CELL NUCLEUS.细胞核中氨基酸的活化及向核糖核酸的转移
Proc Natl Acad Sci U S A. 1959 Oct;45(10):1461-70. doi: 10.1073/pnas.45.10.1461.
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The phosphorus compounds of the cell nucleus.细胞核的磷化合物
Br J Cancer. 1952 Jun;6(2):200-14.
6
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7
STUDIES ON ISOLATED NUCLEI. I. ISOLATION AND CHEMICAL CHARACTERIZATION OF A NUCLEAR FRACTION FROM GUINEA PIG LIVER.豚鼠肝脏细胞核的研究。I. 豚鼠肝脏细胞核组分的分离与化学特性分析
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[On the selection of organic solvents for the obtaining of chloroplasts in non-aqueous media].[关于在非水介质中获取叶绿体时有机溶剂的选择]
Experientia. 1960 Apr 15;16:165-7. doi: 10.1007/BF02157739.
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Evidence for two metabolically distinct types of ribonucleic acid in chromatin and nucleoli.染色质和核仁中两种代谢不同类型核糖核酸的证据。
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本文引用的文献

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Nucleoproteins and the cytological chemistry of Paramecium nuclei.
Proc Soc Exp Biol Med. 1949 Aug;71(4):537-9. doi: 10.3181/00379727-71-17247.
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Variable and constant components of chromosomes.染色体的可变和恒定成分。
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Intracellular distribution of enzymes; the distribution of oxalacetic oxidase activity in rat liver and rat kidney fractions.酶的细胞内分布;草酰乙酸氧化酶活性在大鼠肝脏和大鼠肾脏组分中的分布
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Cytochemical studies of mammalian tissues; the distribution of diphosphopyridine nucleotide-cytochrome c reductase in rat liver fractions.哺乳动物组织的细胞化学研究;大鼠肝脏各组分中辅酶Ⅰ-细胞色素c还原酶的分布
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A study of the behaviour of some 60 amino-acids and other ninhydrin-reacting substances on phenol-collidine filter-paper chromatograms, with notes as to the occurrence of some of them in biological fluids.一项关于约60种氨基酸及其他与茚三酮反应的物质在酚-可力丁滤纸色谱上行为的研究,并记录了其中一些物质在生物体液中的存在情况。
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ISOLATION AND PROPERTIES OF CHICKEN ERYTHROCYTE NUCLEI.鸡红细胞核的分离与特性。
Science. 1943 Jun 25;97(2530):584-5. doi: 10.1126/science.97.2530.584.
7
The amino-aciduria in Fanconi syndrome. A study making extensive use of techniques based on paper partition chromatography.范科尼综合征中的氨基酸尿症。一项大量运用基于纸分配色谱法技术的研究。
Biochem J. 1947;41(2):240-53. doi: 10.1042/bj0410240.
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Qualitative analysis of proteins: a partition chromatographic method using paper.蛋白质的定性分析:一种使用滤纸的分配色谱法。
Biochem J. 1944;38(3):224-32. doi: 10.1042/bj0380224.
9
Antigenic properties of nuclei segregated from spleens of normal and leukemic mice.从正常小鼠和白血病小鼠脾脏中分离出的细胞核的抗原特性。
Cancer Res. 1949 Nov;9(11):669-71.