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用于序列特异性核酸内切酶的生物特异性分级分离基质。

Biospecific fractionation matrices for sequence specific endonucleases.

作者信息

George J, Chirikjian J G

出版信息

Nucleic Acids Res. 1978 Jul;5(7):2223-32. doi: 10.1093/nar/5.7.2223.

Abstract

Fractionation of several type II specific restriction endonucleases was achieved by separation on two novel biospecific matrices. The matrices are pyran, a copolymer of divinyl ether of maleic anhydride, and Cibacron Blue F3GA, a blue dye commonly used for the calibration of molecular sieves. Both compounds are insolubilized by coupling to sepharose through a cyanogen bromide linkage and in their soluble form inhibit the restriction endonucleases which we have tested. These affinity matrices can be used to obtain restriction endonucleases from crude extracts after removal of nucleic acids. They have also proven to have a high capacity when used as subsequent steps in enzyme purification. Their additional advantage is the rapid development time and reusability of columns packed with the two matrices.Images

摘要

通过在两种新型生物特异性基质上进行分离,实现了几种II型特异性限制性内切酶的分级分离。这些基质是吡喃(马来酸酐二乙烯基醚的共聚物)和汽巴克隆蓝F3GA(一种常用于分子筛校准的蓝色染料)。这两种化合物通过溴化氰键与琼脂糖偶联而不溶,并且它们的可溶形式会抑制我们测试过的限制性内切酶。这些亲和基质可用于在去除核酸后从粗提物中获得限制性内切酶。在用作酶纯化的后续步骤时,它们也已证明具有高容量。它们的另一个优点是填充有这两种基质的柱的开发时间短且可重复使用。图像

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/062d/342159/9da79742c644/nar00468-0016-a.jpg

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