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作为杂交探针的单链结合蛋白与M13 DNA复合物。

A complex of single-strand binding protein and M13 DNA as hybridization probe.

作者信息

Syvänen A C, Alanen M, Söderlund H

出版信息

Nucleic Acids Res. 1985 Apr 25;13(8):2789-802. doi: 10.1093/nar/13.8.2789.

Abstract

Single-stranded DNA was complexed to the single-strand binding protein (SSB) of Escherichia coli in a mass ratio of 30:1. The protein moiety of this complex can be labelled by a number of methods of which we have chosen radio-iodination and biotinylation as examples. The SSB-M13 DNA complexes, labelled to high specific activities, were used as probes in hybridization experiments in which 1.6 X 10(-18) moles of immobilized target DNA were detected. The stability of the hybrids was not severely decreased by the binding of SSB. Analysis of hybrids by electron microscopy showed that complexing of DNA with SSB could be used to allow its subsequent identification in the hybrids.

摘要

单链DNA与大肠杆菌的单链结合蛋白(SSB)以30:1的质量比复合。该复合物的蛋白质部分可用多种方法标记,我们选择了放射性碘化和生物素化作为例子。标记至高比活度的SSB - M13 DNA复合物被用作杂交实验中的探针,在该实验中检测到了1.6×10⁻¹⁸摩尔固定化的靶DNA。SSB的结合并未严重降低杂交体的稳定性。通过电子显微镜对杂交体进行分析表明,DNA与SSB的复合可用于在杂交体中对其进行后续鉴定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dffb/341194/d7598b9b964b/nar00302-0124-a.jpg

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