Suppr超能文献

乳糖阻遏蛋白的位点特异性DNA亲和层析

Site-specific DNA-affinity chromatography of the lac repressor.

作者信息

Herrick G

出版信息

Nucleic Acids Res. 1980 Aug 25;8(16):3721-8. doi: 10.1093/nar/8.16.3721.

DOI:10.1093/nar/8.16.3721
PMID:7001362
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC324186/
Abstract

To test the feasibility of site-specific DNA-affinity chromatography, E. coli lac repressor was bound to an operator-containing DNA column, and in parallel to a non-operator DNA column. Salt gradient elution shows: 1) elution from non-operator DNA was near 250mM KCl or NaCl; interpretation of this result suggests the usefulness of the procedure for studying salt-dependence of DNA-protein affinities; 2) elution from operator-containing DNA was delayed (average elution = 1000mM salt), demonstrating a feasibility of site-specific DNA-affinity chromatography, if one provides a sufficiently favorable ratio of specific to non-specific DNA binding sites; 3) repressor eluted from operator-containing DNA over a very broad salt range, which may represent chromatography-generated repressor heterogeneity.

摘要

为测试位点特异性DNA亲和色谱的可行性，将大肠杆菌乳糖阻遏物与含操纵基因的DNA柱结合，并同时与不含操纵基因的DNA柱结合。盐梯度洗脱显示：1）从不含操纵基因的DNA上洗脱接近250mM KCl或NaCl；对该结果的解释表明该方法对于研究DNA-蛋白质亲和力的盐依赖性很有用；2）从含操纵基因的DNA上洗脱延迟（平均洗脱盐浓度 = 1000mM），这表明如果提供足够有利的特异性与非特异性DNA结合位点比例，位点特异性DNA亲和色谱是可行的；3）阻遏物从含操纵基因的DNA上在非常宽的盐浓度范围内洗脱，这可能代表色谱产生的阻遏物异质性。

相似文献

1
Site-specific DNA-affinity chromatography of the lac repressor.乳糖阻遏蛋白的位点特异性DNA亲和层析
Nucleic Acids Res. 1980 Aug 25;8(16):3721-8. doi: 10.1093/nar/8.16.3721.
2
Isolation of DNA-protein complexes based on streptavidin and biotin interaction.
Eur J Biochem. 1987 Jul 15;166(2):351-5. doi: 10.1111/j.1432-1033.1987.tb13522.x.
3
Heparin elution of transcription factors from DNA-Sepharose columns.从DNA-琼脂糖柱上洗脱转录因子的肝素法
J Chromatogr A. 1999 Jul 2;848(1-2):131-8. doi: 10.1016/s0021-9673(99)00338-6.
4
Temperature dependence of DNA affinity chromatography of transcription factors.转录因子的DNA亲和色谱法的温度依赖性
Anal Biochem. 2000 Mar 15;279(2):209-17. doi: 10.1006/abio.2000.4489.
5
Purification of Lac repressor protein using polymer displacement and immobilization of the protein.
Bioseparation. 1999;8(6):307-16. doi: 10.1023/a:1008101711218.
6
Gel retardation at low pH resolves trp repressor-DNA complexes for quantitative study.在低pH值下的凝胶阻滞分析可分离色氨酸阻遏物 - DNA复合物用于定量研究。
Proc Natl Acad Sci U S A. 1988 Feb;85(4):975-9. doi: 10.1073/pnas.85.4.975.
7
Activity changes in lac repressor with cysteine oxidation.
J Biol Chem. 1979 May 10;254(9):3341-7.
8
NMR study of the interaction between the lac repressor and the lac operator.乳糖阻遏物与乳糖操纵基因相互作用的核磁共振研究
Eur J Biochem. 1983 May 2;132(2):321-7. doi: 10.1111/j.1432-1033.1983.tb07365.x.
9
Escherichia coli lac repressor-lac operator interaction and the influence of allosteric effectors.大肠杆菌乳糖阻遏物-乳糖操纵基因相互作用及别构效应剂的影响
J Mol Biol. 1997 Jan 10;265(1):1-7. doi: 10.1006/jmbi.1996.0706.
10
Comparative studies on chemically and enzymatically coupled DNA-Sepharose columns for purification of a lac repressor chimeric fusion protein.用于纯化乳糖阻遏物嵌合融合蛋白的化学偶联和酶促偶联DNA-琼脂糖柱的比较研究
J Chromatogr A. 1999 Jul 23;849(2):403-12. doi: 10.1016/s0021-9673(99)00586-5.

引用本文的文献

1
Affinity purification of sequence-specific DNA binding proteins.序列特异性DNA结合蛋白的亲和纯化
Proc Natl Acad Sci U S A. 1986 Aug;83(16):5889-93. doi: 10.1073/pnas.83.16.5889.
2
Purification of the c-fos enhancer-binding protein.c-fos增强子结合蛋白的纯化
Mol Cell Biol. 1987 Oct;7(10):3482-9. doi: 10.1128/mcb.7.10.3482-3489.1987.
3
Identification and purification of a polypeptide that binds to the c-fos serum response element.一种与c-fos血清反应元件结合的多肽的鉴定与纯化。
EMBO J. 1987 Sep;6(9):2711-7. doi: 10.1002/j.1460-2075.1987.tb02564.x.
4
A high-capacity column for affinity purification of sequence-specific DNA-binding proteins.用于序列特异性DNA结合蛋白亲和纯化的高容量柱。
Nucleic Acids Res. 1992 Jul 11;20(13):3525. doi: 10.1093/nar/20.13.3525.

本文引用的文献

1
A deoxyribonucleic acid polymerase from Micrococcus luteus (Micrococcus lysodeikticus) isolated on deoxyribonucleic acid-cellulose.从微球菌（溶壁微球菌）中分离得到的一种脱氧核糖核酸聚合酶，该酶是在脱氧核糖核酸 - 纤维素上分离得到的。
J Biol Chem. 1968 Dec 10;243(23):6222-33.
2
The lac repressor-operator interaction. 3. Kinetic studies.乳糖阻遏蛋白与操纵基因的相互作用。3. 动力学研究。
J Mol Biol. 1970 Nov 14;53(3):401-17. doi: 10.1016/0022-2836(70)90074-4.
3
Lac repressor-operator interaction. I. Equilibrium studies.乳糖阻遏蛋白与操纵基因的相互作用。I. 平衡研究。
J Mol Biol. 1970 Feb 28;48(1):67-83. doi: 10.1016/0022-2836(70)90219-6.
4
Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form.大肠杆菌中的超螺旋环状DNA-蛋白质复合物：纯化及诱导转化为开放环状DNA形式
Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159-66. doi: 10.1073/pnas.62.4.1159.
5
Plasmid ColEl as a molecular vehicle for cloning and amplification of DNA.质粒ColE1作为用于DNA克隆和扩增的分子载体。
Proc Natl Acad Sci U S A. 1974 Sep;71(9):3455-9. doi: 10.1073/pnas.71.9.3455.
6
The location of the repressor binding sites in the lac operon.阻遏蛋白结合位点在乳糖操纵子中的位置。
Proc Natl Acad Sci U S A. 1974 Jun;71(6):2314-8. doi: 10.1073/pnas.71.6.2314.
7
lac Repressor-operator interaction. IX. The binding of lac repressor to operators containing Oc mutations.乳糖阻遏蛋白-操纵基因相互作用。IX. 乳糖阻遏蛋白与含Oc突变的操纵基因的结合。
J Mol Biol. 1974 May 15;85(2):231-48. doi: 10.1016/0022-2836(74)90362-3.
8
Lac repressor binding to non-operator DNA: detailed studies and a comparison of eequilibrium and rate competition methods.乳糖阻遏蛋白与非操纵基因DNA的结合：详细研究以及平衡和速率竞争方法的比较
J Mol Biol. 1972 Dec 30;72(3):671-90. doi: 10.1016/0022-2836(72)90184-2.
9
Lac repressor binding to poly (d(A-T)). Conformational changes.乳糖阻遏蛋白与聚（d（A-T））的结合。构象变化。
Biochem Biophys Res Commun. 1974 Oct 8;60(3):951-7. doi: 10.1016/0006-291x(74)90406-9.
10
Theoretical aspects of DNA-protein interactions: co-operative and non-co-operative binding of large ligands to a one-dimensional homogeneous lattice.DNA-蛋白质相互作用的理论方面：大配体与一维均匀晶格的协同和非协同结合。
J Mol Biol. 1974 Jun 25;86(2):469-89. doi: 10.1016/0022-2836(74)90031-x.

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验