Krämer A, Bautz E K
Eur J Biochem. 1981 Jul;117(3):449-55. doi: 10.1111/j.1432-1033.1981.tb06359.x.
A procedure is described which allows rapid purification of RNA polymerase II from either embryos of Drosophila melanogaster or larvae of Chironomus thummi. The polypeptide compositions of the enzymes were compared in denaturing gels. Antisera were raised against the native enzymes and were tested for binding activity to separated subunits of each enzyme. Sheep antibodies to Drosophila RNA polymerase were found to react with most polypeptides of the insect RNA polymerases but only with the high-molecular weight subunits of calf thymus RNA polymerase II. These were also found to cross-react with a monoclonal antibody directed against the large subunits of the Drosophila enzyme. An antiserum to Chironomus RNA polymerase II reacted with the three largest subunits of this enzyme and subunits 0, 1 and 2 of Drosophila RNA polymerase II. The purified IgG fractions of the polyspecific antisera inhibited the activity in vitro of the insect RNA polymerases to different degrees.
本文描述了一种可从黑腹果蝇胚胎或图氏摇蚊幼虫中快速纯化RNA聚合酶II的方法。在变性凝胶中比较了这些酶的多肽组成。制备了针对天然酶的抗血清，并检测其与每种酶分离亚基的结合活性。发现针对果蝇RNA聚合酶的羊抗体可与昆虫RNA聚合酶的大多数多肽发生反应，但仅与小牛胸腺RNA聚合酶II的高分子量亚基发生反应。还发现这些抗体与针对果蝇酶大亚基的单克隆抗体发生交叉反应。针对摇蚊RNA聚合酶II的抗血清与该酶的三个最大亚基以及果蝇RNA聚合酶II的0、1和2亚基发生反应。多特异性抗血清的纯化IgG组分在不同程度上抑制了昆虫RNA聚合酶的体外活性。
