摘要
South-Westerm blot mapping是一种结合Western blotting和Southern blotting某些特点的方法.本文介绍用其成功地观察到锥虫核蛋白中DNA结合蛋白的情况,并对一个分子量在40000左右、于较严谨条件下与DNA结合的核蛋白进行了特性鉴定.该蛋白等量地存在于锥虫的前循环期和血液期,对双链DNA有较大的亲和力,并能与酵母菌复制起始片段结合.本文还介绍了锥虫细胞核的提取技术和核蛋白的制备技术.
A method which is referred to as 'South-Western blotting mapping' is described. Nuclear extracts of trypanosomes are separated on SDS-polyacrylamide gel, blotted onto nitrocellulose by electrotransfer and hybridized with end-labelled DNA fragments. DNA-binding protein (proteins) is further characterized. The protein with Mr of 40000 is present in both blood stream and procyclic parasites. It has preferential affinity for the double-stranded DNA and binds yeast replication origin fragments at the same intensity as those of trypanosome genomic DNA. A method for preparation of nuclear proteins is also described.
关键词
DNA
结合蛋白
基因组
锥虫
South-Western blotting mapping
DNA-binding protein
genomic DNA
nu- clear proteins
trypanosomes
