摘要
本文运用凝胶延迟反应（ＧｅｌＲｅｔａｒｄａｔｉｏｎＡｓｓａｙ）和足迹法（Ｆｏｏｔｐｒｉｎｔｉｎｇ）研究了ＨｅＬａ细胞核蛋白中的反式作用因子ＣＴＦ／ＮＦ－１与其对应的顺式作用元件的特异性作用。研究结果表明ＨｅＬａ细胞的ＣＴＦ／ＮＦ－１包含有不同分子量的组分，它们均能特异性结合于特异位点ＡＴＡＴＴＧＧＣＴＴＣＡＡＧＣＣＡＡＡＡＴＧＧＡ序列，形成至少５种不同分子量形式的因子－元件复合物。这一结果对阐明ＣＴＦ／ＮＦ－１复杂多样的调控机制及其组成种类有重要意义。此外，本文还建立了一种制备核蛋白提取物的新方法，有效地解决了因反式作用因子含量极低带来的研究上的困难。
The specific reaction between the trans-acting factor,CTF/NF-1, and the related cis-acting element was studied. On gel retardation assay and footprinting analysis. it was demonstrated that there were at least five composed elements of CTF/NF-1 in the nuclear extracts of HeLa cells. All of them can specifically bind to 0the DNA sequence ATATTGGCTTCAAGCCAAAATGGA in vitro, As a result. at least five specific compounds of the protein-nuclear acid were formed. These results can help us to learn the composition of CTF/NF-1 and explain the complicated functional mechanism of CTF/NF-1 in the process of DNA replication and transcription. In this paper, we also built a new method used for preparing the nuclear extracts. Consequently, the problem was resolved which is caused by very slight quantity of the factors contained in eukaryotic cells.
基金
中国科学院遗传研究所所长基金
关键词
反式作用因子
顺式作用
HELA细胞
CTF/NF-1, Trans-acting factor, Cis-acting element, Gel retardation assay, Footprinting
