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从大鼠肝细胞含内质网亚组分中制备聚（A）结合蛋白及其在mRNA纯化中的应用。

Preparation of poly (A)-binding proteins from endoplasmic reticulum-containing subfractions of rat liver cells and their use in mRNA purification.

作者信息

Axelsson B, Ohlsson R, Deutsch A, Jergil B

出版信息

Mol Cell Biochem. 1977 Mar 21;15(1):67-71. doi: 10.1007/BF01731290.

DOI:10.1007/BF01731290
PMID:865485
Abstract

Approximately 2% of the proteins solubilised from rat liver microsomes or rapidly sedimenting endoplasmic reticulum (RS-ER) adsorbed to poly(A)-Sepharose. The adsorption appeared to be selective for a few proteins, and proteins of different apparent molecular weights adsorbed from RS-ER and the microsomes. The proteins from RS-ER with affinity for poly(A) were coupled to Sepharose and used for the purification of mRNA from rabbit mammary glands. A portion of the RNA wihich did not adsorb to poly(U)-Sepharose adsorbed to protein-Sepharose and was active in a cell-free protein synthesis system.

摘要

从大鼠肝脏微粒体或快速沉降内质网（RS-ER）中溶解出来的蛋白质，约有2%吸附到聚（A）-琼脂糖上。这种吸附似乎对少数蛋白质具有选择性，并且从RS-ER和微粒体中吸附的蛋白质具有不同的表观分子量。具有聚（A）亲和力的RS-ER中的蛋白质与琼脂糖偶联，并用于从兔乳腺中纯化mRNA。一部分未吸附到聚（U）-琼脂糖上的RNA吸附到蛋白质-琼脂糖上，并在无细胞蛋白质合成系统中具有活性。

相似文献

1
Preparation of poly (A)-binding proteins from endoplasmic reticulum-containing subfractions of rat liver cells and their use in mRNA purification.从大鼠肝细胞含内质网亚组分中制备聚（A）结合蛋白及其在mRNA纯化中的应用。
Mol Cell Biochem. 1977 Mar 21;15(1):67-71. doi: 10.1007/BF01731290.
2
Segregation of the polypeptide translocation apparatus to regions of the endoplasmic reticulum containing ribophorins and ribosomes. I. Functional tests on rat liver microsomal subfractions.多肽转运装置定位于内质网中含有核糖体结合蛋白和核糖体的区域。I. 对大鼠肝脏微粒体亚组分的功能测试。
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3
In vitro translation of globin: effect of proteins purified by affinity chromatography on polyadenylate-Sepharose.珠蛋白的体外翻译：通过聚腺苷酸-琼脂糖亲和层析纯化的蛋白质的作用。
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Eur J Biochem. 1977 Feb;72(3):595-603. doi: 10.1111/j.1432-1033.1977.tb11282.x.

本文引用的文献

1
Protein measurement with the Folin phenol reagent.使用福林酚试剂进行蛋白质测定。
J Biol Chem. 1951 Nov;193(1):265-75.
2
ISOLATION AND CHARACTERIZATION OF RIBOSOMAL RIBONUCLEIC ACID.核糖体核糖核酸的分离与鉴定
Biochem J. 1965 Jul;96(1):266-9. doi: 10.1042/bj0960266.
3
"Molecular sieve" chromatography on polyacrylamide gels, prepared according to a simplified method.根据简化方法制备的聚丙烯酰胺凝胶上的“分子筛”色谱法。
Arch Biochem Biophys. 1962 Sep;Suppl 1:147-51.
4
Chemical coupling of peptides and proteins to polysaccharides by means of cyanogen halides.通过卤化氰将肽和蛋白质与多糖进行化学偶联。
Nature. 1967 Jun 24;214(5095):1302-4. doi: 10.1038/2141302a0.
5
An adenylic acid-rich sequence in messenger RNA of HeLa cells and its possible relationship to reiterated sites in DNA.海拉细胞信使核糖核酸中富含腺苷酸的序列及其与DNA中重复位点的可能关系。
Proc Natl Acad Sci U S A. 1971 Jun;68(6):1321-5. doi: 10.1073/pnas.68.6.1321.
6
Preparation of sepharose-bound poly (rI:rC).琼脂糖结合的聚（rI：rC）的制备。
Biochem Biophys Res Commun. 1971 Oct 1;45(1):184-9. doi: 10.1016/0006-291x(71)90067-2.
7
Molecular weight determination of protein-dodecyl sulfate complexes by gel electrophoresis in a discontinuous buffer system.在不连续缓冲系统中通过凝胶电泳测定蛋白质 - 十二烷基硫酸盐复合物的分子量
J Biol Chem. 1971 Oct 25;246(20):6328-34.
8
Isolation of mRNA from KB-cells by affinity chromatography on polyuridylic acid covalently linked to Sepharose.通过在共价连接到琼脂糖凝胶的聚尿苷酸上进行亲和层析从KB细胞中分离信使核糖核酸。
Eur J Biochem. 1972 Dec 4;31(2):246-54. doi: 10.1111/j.1432-1033.1972.tb02527.x.
9
Characterization of a protein species isolated from HeLa cell cytoplasm by affinity chromatography on polyadenylate-sepharose.通过聚腺苷酸-琼脂糖亲和层析从人宫颈癌（HeLa）细胞胞质中分离出的一种蛋白质的特性分析。
Proc Natl Acad Sci U S A. 1974 May;71(5):1882-6. doi: 10.1073/pnas.71.5.1882.
10
Efficient translation of tobacco mosaic virus RNA and rabbit globin 9S RNA in a cell-free system from commercial wheat germ.在由市售小麦胚芽制备的无细胞体系中烟草花叶病毒RNA和兔珠蛋白9S RNA的高效翻译。
Proc Natl Acad Sci U S A. 1973 Aug;70(8):2330-4. doi: 10.1073/pnas.70.8.2330.