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新DNA结合蛋白的系留条带迁移分析及亲和纯化

Tethered bandshift assay and affinity purification of a new DNA-binding protein.

作者信息

Ren L, Chen H, Sternberg E A

机构信息

Medical College of Wisconsin, Milwaukee.

出版信息

Biotechniques. 1994 May;16(5):852-5.

PMID:8068340
Abstract

DNA-binding proteins can be purified by their affinity for probes with specific sequences that are tethered to magnetic beads. A restriction site at the end of the tether allows release of probe with factor still on it. This probe with bound factor can be bandshifted directly to compare to bandshifts in whole extracts. This method correlates purified factor seen indirectly by bandshift with factor that is purified and released, because they are the same.

摘要

DNA结合蛋白可以通过其对与固定在磁珠上的特定序列探针的亲和力来纯化。连接物末端的限制酶切位点允许仍带有因子的探针被释放。这种带有结合因子的探针可以直接进行凝胶迁移分析,以便与全细胞提取物中的凝胶迁移分析结果进行比较。这种方法将通过凝胶迁移间接观察到的纯化因子与纯化并释放的因子联系起来,因为它们是相同的。

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