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[与免疫球蛋白基因八聚体序列相互作用的蛋白质因子]

[Protein factors, interacting with the octamer sequence of immunoglobulin genes].

作者信息

Stepchenko A G, Luchina N N, Polianovskiĭ O L

出版信息

Mol Biol (Mosk). 1989 May-Jun;23(3):708-16.

PMID:2770740
Abstract

Nuclei of different cell lines contain protein factors interacting with octamer ATTTGCAT. Fragment k53 of kappa-gene promoter region was used as DNA-probe. The factors from lymphoid cells yield a DNA/protein complex with mobility B0. The proteins are referred to as HF-B0. The nonspecific ubiquitous factor present in many non-lymphoid cells (for instance, HeLa cells) interacts with the probe to produce a complex whose mobility is much lower. The protein NF-B0 was isolated from the nuclear extract of myeloma MOPC21 cells. It was purified by chromatography on ion exchangers, hydroxylapatite, heparin-Sepharose and affinity sorbent containing a synthetic octamer sequence. At all the steps of purification, protein fractions were chosen for their ability to interact selectively with the octamer yielding a complex with the mobility B0. As a result, NF-B0 protein (60 +/- 2)kDa was purified 6.10(4) times to the electrophoretically homogeneous state. Purified factor NF-B0 selectively interacts with the octamer.

摘要

不同细胞系的细胞核含有与八聚体ATTTGCAT相互作用的蛋白质因子。κ基因启动子区域的片段k53用作DNA探针。来自淋巴细胞的因子产生迁移率为B0的DNA/蛋白质复合物。这些蛋白质被称为HF-B0。许多非淋巴细胞(例如,HeLa细胞)中存在的非特异性普遍因子与探针相互作用,产生迁移率低得多的复合物。蛋白质NF-B0是从骨髓瘤MOPC21细胞的核提取物中分离出来的。它通过在离子交换剂、羟基磷灰石、肝素-琼脂糖和含有合成八聚体序列的亲和吸附剂上进行色谱法纯化。在纯化的所有步骤中,根据蛋白质组分与八聚体选择性相互作用产生迁移率为B0的复合物的能力来选择。结果,将(60±2)kDa的NF-B0蛋白纯化了6.10⁴倍,达到电泳纯状态。纯化的因子NF-B0与八聚体选择性相互作用。

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