汞化RNA在巯基琼脂糖色谱法中的RNA聚集
RNA aggregation during sulfhydryl-agarose chromatography of mercurated RNA.
作者信息
Konkel D A, Ingram V M
出版信息
Nucleic Acids Res. 1977 Jun;4(6):1979-88. doi: 10.1093/nar/4.6.1979.
Abstract
Isolation of newly synthesized mercurated RNA transcripts by chromatography on sulfhydryl-agarose has recently been used to reduce contamination by endogenous RNA derived from the chromatin template. We show that substantial RNA aggregation occurs during standard isolation procedures, causing significant retention of endogenous (unmercurated) RNA on sulfhydryl-agarose. We describe methods to reduce substantially this problem and discuss the implications of our findings for interpretation of previous hybridization and transcription experiments.
摘要
最近，通过巯基琼脂糖层析分离新合成的汞化RNA转录本已被用于减少来自染色质模板的内源性RNA的污染。我们发现，在标准分离过程中会发生大量RNA聚集，导致内源性（未汞化）RNA在巯基琼脂糖上大量保留。我们描述了大幅减少这一问题的方法，并讨论了我们的发现对解释先前杂交和转录实验的影响。
相似文献
1
RNA aggregation during sulfhydryl-agarose chromatography of mercurated RNA.汞化RNA在巯基琼脂糖色谱法中的RNA聚集
Nucleic Acids Res. 1977 Jun;4(6):1979-88. doi: 10.1093/nar/4.6.1979.
2
In vitro transcription of chromatin in the presence of a mercurated nucleotide.在含有汞化核苷酸的情况下对染色质进行体外转录。
Proc Natl Acad Sci U S A. 1976 May;73(5):1564-7. doi: 10.1073/pnas.73.5.1564.
3
A method for purification of mercurated nucleic acid hybrids on ether-linked sulfhydryl-agarose.一种在醚连接的巯基琼脂糖上纯化汞化核酸杂交体的方法。
Anal Biochem. 1978 Sep;89(2):501-7. doi: 10.1016/0003-2697(78)90379-2.
4
Is there specific transcription from isolated chromatin?从分离的染色质中是否存在特异性转录？
Nucleic Acids Res. 1978 Apr;5(4):1237-52. doi: 10.1093/nar/5.4.1237.
5
Purification of DNA or RNA sequences complementary to mercurated recombinant DNA by sulfhydryl sepharose chromatography.
Methods Enzymol. 1979;68:192-9. doi: 10.1016/0076-6879(79)68014-x.
6
In vitro DNA dependent synthesis of globin RNA sequences from erythroleukemic cell chromatin.从红白血病细胞染色质体外依赖DNA合成珠蛋白RNA序列。
Nucleic Acids Res. 1979 Jan;6(1):275-87. doi: 10.1093/nar/6.1.275.
7
Isolation of globin pre-messenger RNA on thiol-agarose by terminally mercurated complementary DNA.通过末端汞化互补DNA在硫醇琼脂糖上分离珠蛋白前体信使RNA。
Mol Biol Rep. 1981 May 22;7(1-3):83-92. doi: 10.1007/BF00778737.
8
Estrogen withdrawal in chick oviduct. Characterization of RNA synthesized in isolated nuclei using a mercurated precursor.雏鸡输卵管中的雌激素撤退。使用汞化前体对分离细胞核中合成的RNA进行表征。
Biochim Biophys Acta. 1978 Aug 23;520(1):184-202. doi: 10.1016/0005-2787(78)90019-9.
9
Fidelity of chromatin transcription in vitro.体外染色质转录的保真度。
Biochemistry. 1976 Oct 5;15(20):4356-63. doi: 10.1021/bi00665a002.
10
Mercurated polynucleotides: new probes for hybridization and selective polymer fractionation.汞化多核苷酸：用于杂交和选择性聚合物分级分离的新型探针。
Biochemistry. 1975 Jun 3;14(11):2458-69. doi: 10.1021/bi00682a028.
引用本文的文献
1
Prolactin induces release of a factor from membranes capable of stimulating beta-casein gene transcription in isolated mammary cell nuclei.催乳素可诱导细胞膜释放一种因子，该因子能够刺激分离出的乳腺细胞核中β-酪蛋白基因的转录。
Proc Natl Acad Sci U S A. 1981 Nov;78(11):6729-33. doi: 10.1073/pnas.78.11.6729.
2
Characterization of the 5' termini of purified nascent simian virus 40 late transcripts.纯化的猿猴病毒40晚期转录本5'末端的特征分析。
J Virol. 1983 Jan;45(1):264-74. doi: 10.1128/JVI.45.1.264-274.1983.
3
Simian virus 40 transcriptional complexes incorporate mercurated nucleotides into RNA in vitro.猿猴病毒40转录复合物在体外将汞化核苷酸掺入RNA中。
J Virol. 1981 Jan;37(1):511-7. doi: 10.1128/JVI.37.1.511-517.1981.
4
Characterization of RNA synthesized in isolated nuclei of herpes simplex virus type 1-infected KB cells.单纯疱疹病毒1型感染的KB细胞核中合成的RNA的特性分析。
J Virol. 1980 Jan;33(1):567-71. doi: 10.1128/JVI.33.1.567-571.1980.
5
Hormonal modulation of alpha-fetoprotein gene expression in newborn rat livers.新生大鼠肝脏中甲胎蛋白基因表达的激素调节
Nucleic Acids Res. 1981 Dec 21;9(24):6917-33. doi: 10.1093/nar/9.24.6917.
6
A new hybridocytochemical method based on mercurated nucleic acid probes and sulfhydryl-hapten ligands. I. Stability of the mercury-sulfhydryl bond and influence of the ligand structure on immunochemical detection of the hapten.一种基于汞化核酸探针和巯基半抗原配体的新型杂交细胞化学方法。I. 汞-巯基键的稳定性及配体结构对半抗原免疫化学检测的影响。
Histochemistry. 1986;84(2):169-78. doi: 10.1007/BF00499829.
7
The reactions of mercurated pyrimidine nucleotides with thiols and with hydrogen sulfide.汞化嘧啶核苷酸与硫醇及硫化氢的反应。
Nucleic Acids Res. 1978 Jun;5(6):2133-51. doi: 10.1093/nar/5.6.2133.
8
Is there specific transcription from isolated chromatin?从分离的染色质中是否存在特异性转录？
Nucleic Acids Res. 1978 Apr;5(4):1237-52. doi: 10.1093/nar/5.4.1237.
9
Affinity chromatography of thiol-containing purines and ribonucleic acid.含硫醇嘌呤和核糖核酸的亲和层析
Biochem J. 1977 Dec 15;168(3):595-7. doi: 10.1042/bj1680595.
10
Synthesis and processing of adenoviral RNA in isolated nuclei.腺病毒RNA在分离细胞核中的合成与加工
J Virol. 1979 Nov;32(2):394-403. doi: 10.1128/JVI.32.2.394-403.1979.
本文引用的文献
1
Tissue sulfhydryl groups.组织巯基
Arch Biochem Biophys. 1959 May;82(1):70-7. doi: 10.1016/0003-9861(59)90090-6.
2
Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads.通过亲和色谱法进行蛋白质纯化。琼脂糖和聚丙烯酰胺珠粒的衍生化。
J Biol Chem. 1970 Jun;245(12):3059-65.
3
A new method for the large scale purification of Escherichia coli deoxyribonucleic acid-dependent ribonucleic acid polymerase.一种大规模纯化大肠杆菌脱氧核糖核酸依赖性核糖核酸聚合酶的新方法。
J Biol Chem. 1969 Nov 25;244(22):6160-7.
4
Preparation of chromatin. Variation in the template properties of chromatin dependent on the method of perparation.染色质的制备。染色质模板特性的变化取决于制备方法。
Eur J Biochem. 1974 Aug 1;46(3):461-71. doi: 10.1111/j.1432-1033.1974.tb03639.x.
5
Histones in chick embryonic erythropoiesis.鸡胚胎红细胞生成中的组蛋白。
J Biol Chem. 1973 Feb 10;248(3):1025-31.
6
Tissue-specific transcription of the globin gene in isolated chromatin.分离染色质中珠蛋白基因的组织特异性转录。
Proc Natl Acad Sci U S A. 1973 Dec;70(12):3440-2. doi: 10.1073/pnas.70.12.3440.
7
Synthesis of globin ribonucleic acid from duck-reticulocyte chromatin in vitro.体外从鸭网织红细胞染色质合成珠蛋白核糖核酸。
Proc Natl Acad Sci U S A. 1973 Jul;70(7):2029-32. doi: 10.1073/pnas.70.7.2029.
8
Purification and properties of biologically active messenger RNA for a myeloma light chain.骨髓瘤轻链生物活性信使核糖核酸的纯化及特性
Proc Natl Acad Sci U S A. 1972 Jul;69(7):1967-71. doi: 10.1073/pnas.69.7.1967.
9
Lineage-dependent transcription of globin genes.珠蛋白基因的谱系依赖性转录。
Cell. 1974 Nov;3(3):243-7. doi: 10.1016/0092-8674(74)90138-x.
10
Regulation of specific gene expression during embryonic development: Synthesis of globin messenger RNA during red cell formation in chick embryos.胚胎发育过程中特定基因表达的调控：鸡胚红细胞形成过程中珠蛋白信使核糖核酸的合成。
Dev Biol. 1974 Sep;40(1):174-85. doi: 10.1016/0012-1606(74)90117-1.
Zotero 插件