Oligonucleotide-directed nucleic acid scission by micrococcal nuclease.
Oligonucleotide-directed nucleic acid scission by micrococcal nuclease.
复制标题
通过微球菌核酸酶进行寡核苷酸指导的核酸切割。
DOI：
10.1021/bi00201a006
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发表时间：
1994
期刊：
影响因子：
2.9
通讯作者：
Sigman,DS
中科院分区：
文献类型：
--
作者：
Landgraf,R;Chen,CH;Sigman,DS
19948年5月17日收到的修订手稿摘要：通过使用Cibacron Blue(CB)F3G-A将由谷胱甘肽5-转移酶和减毒微球菌核酸酶(MN)组成的融合蛋白与靶向寡核苷酸捆绑在一起，可以实现由序列决定的dna切割。在凝胶迁移率改变分析中，用该染料衍生的脱氧寡核苷酸与融合蛋白结合。这种结合方案被成功地用于与CB衍生的互补寡核苷酸杂交后实现单链DNA底物的定点切割。尽管过去已经成功地使用MN和寡核苷酸的共价交联杂交物来靶向核溶解活性，但这种新的方案为靶向和探查两者打开了新的可能性
Revised Manuscript Received May 17, 19948 abstract:“Sequence-dictated” scission of DNA can be achieved by tethering a fusion protein composed of glutathione 5-transferase and attenuated micrococcal nuclease (MN) to a targeting oligonucleotide using Cibacron blue (CB) F3G-A. Deoxyoligonucleotides derivatized with this dye bind to the fusionprotein in gel mobility shift assays. This binding scheme was successfully used to achieve site-specific scission of a single-stranded DNA substrate after hybridization with a CB-derivatized complementary oligonucleotide. Although covalently cross-linked hybrids of MN and oligonucleotides have been successfully used in the past to target nucleolytic activity, this novel scheme opens new possibilities for targeting and probing both