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抗生物素蛋白-生物素基因富集方法在含有特定基因序列的长双链DNA分离中的应用。

Application of the avidin-biotin method of gene enrichment to the isolation of long double-stranded DNA containing specific gene sequences.

作者信息

Pellegrini M, Holmes D S, Manning J

出版信息

Nucleic Acids Res. 1977 Sep;4(9):2961-73. doi: 10.1093/nar/4.9.2961.

DOI:10.1093/nar/4.9.2961
PMID:198749
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC342627/
Abstract

A method of enriching for long double-stranded segments of eukaryotic DNA carrying particular genes is described. A purified RNA coded for by the gene is covalently attached to biotin via the protein, cytochrome c. This modified RNA is hybridized to total nuclear, double-stranded DNA under conditions that allow the formation of R-loops. Avidin, which has a high affinity for biotin, is covalently attached to polymer spheres. The complexes of avidin-spheres with DNA:RNA-biotin R-loop hybrids band in CsCl at a much lower bouyant density than does free DNA. This density is a function of the length of DNA coupled per avidin-sphere. This method was used to prepare very long double-strands of DNA highly enriched in the coding sequences for the large rRNAs of D. melanogaster and L. donovani and the histone mRNAs of S. purpuratus.

摘要

本文描述了一种富集携带特定基因的真核生物DNA长双链片段的方法。由该基因编码的纯化RNA通过细胞色素c蛋白与生物素共价连接。在允许形成R环的条件下，这种修饰的RNA与总的核双链DNA杂交。对生物素有高亲和力的抗生物素蛋白与聚合物球体共价连接。抗生物素蛋白-球体与DNA:RNA-生物素R环杂交体的复合物在CsCl中的浮力密度比游离DNA低得多。这种密度是每个抗生物素蛋白-球体结合的DNA长度的函数。该方法用于制备高度富集黑腹果蝇和杜氏利什曼原虫大rRNA编码序列以及紫球海胆组蛋白mRNA的非常长的双链DNA。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/776b/342627/e290c0a74020/nar00482-0029-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/776b/342627/133edac0a016/nar00482-0025-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/776b/342627/e290c0a74020/nar00482-0029-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/776b/342627/133edac0a016/nar00482-0025-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/776b/342627/e290c0a74020/nar00482-0029-a.jpg

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本文引用的文献

1
EQUILIBRIUM SEDIMENTATION OF MACROMOLECULES IN DENSITY GRADIENTS.密度梯度中大分子的平衡沉降
Proc Natl Acad Sci U S A. 1957 Jul 15;43(7):581-8. doi: 10.1073/pnas.43.7.581.
2
Gene linkage by RNA-DNA hybridization. I. Unique DNA sequences homologous to 4 s RNA, 5 s RNA and ribosomal RNA.通过RNA-DNA杂交进行基因连锁分析。I. 与4s RNA、5s RNA和核糖体RNA同源的独特DNA序列。
J Mol Biol. 1968 Jun 28;34(3):661-80. doi: 10.1016/0022-2836(68)90188-5.
3
Properties and composition of the isolated ribosomal DNA satellite of Xenopus laevis.
Nucleic Acids Res. 1978 Feb;5(2):491-504. doi: 10.1093/nar/5.2.491.
4
Electron microscopic visualization of tRNA genes with ferritin-avidin: biotin labels.用铁蛋白抗生物素蛋白：生物素标记对转运RNA基因进行电子显微镜观察。
Nucleic Acids Res. 1978 Feb;5(2):363-84. doi: 10.1093/nar/5.2.363.
5
Strand-specific attachment of avidin-spheres to double-stranded poliovirus RNA.
Proc Natl Acad Sci U S A. 1979 Feb;76(2):676-80. doi: 10.1073/pnas.76.2.676.
6
Gene mapping and gene enrichment by the avidin-biotin interaction: use of cytochrome-c as a polyamine bridge.
Nucleic Acids Res. 1978 Feb;5(2):385-401. doi: 10.1093/nar/5.2.385.
非洲爪蟾分离核糖体DNA卫星的性质与组成
Nature. 1968 Aug 3;219(5153):454-63. doi: 10.1038/219454a0.
4
Hybridization of filters with competitor DNA in the liquid phase in a standard and a micro-assay.在标准检测和微量检测中，滤膜与竞争DNA在液相中的杂交。
Biochimie. 1974;56(9):1215-21. doi: 10.1016/s0300-9084(74)80014-3.
5
Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli. I. Structure of F-prime factors.大肠杆菌质粒间序列关系的电子显微镜异源双链研究。I. F-prime因子的结构
J Mol Biol. 1972 Nov 14;71(2):471-97. doi: 10.1016/0022-2836(72)90363-4.
6
Detection of specific sequences among DNA fragments separated by gel electrophoresis.在通过凝胶电泳分离的DNA片段中检测特定序列。
J Mol Biol. 1975 Nov 5;98(3):503-17. doi: 10.1016/s0022-2836(75)80083-0.
7
Physicochemical properties of kinetoplast DNA from Crithidia acanthocephali. Crithidia luciliae, and Trypanosoma lewisi.来自棘头克氏锥虫、鲁氏克氏锥虫和路氏锥虫的动基体DNA的物理化学性质。
J Cell Biol. 1975 Nov;67(2PT.1):378-99. doi: 10.1083/jcb.67.2.378.
8
Isolation of histone genes from unfractionated sea urchin DNA by subculture cloning in E. coli.通过在大肠杆菌中进行亚克隆培养从未分级的海胆DNA中分离组蛋白基因。
Nature. 1975 Jun 12;255(5509):533-8. doi: 10.1038/255533a0.
9
The organization of sea urchin histone genes.海胆组蛋白基因的组织方式。
Cell. 1975 Nov;6(3):359-69. doi: 10.1016/0092-8674(75)90185-3.
10
Determination of DNA concentration by electron microscopy.通过电子显微镜测定DNA浓度。
Anal Biochem. 1976 May 7;72:460-7. doi: 10.1016/0003-2697(76)90554-6.