Cho H, Maldonado E, Reinberg D
Howard Hughes Medical Institute, Division of Nucleic Acid Enzymology, Department of Biochemistry, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway, New Jersey 08854, USA.
J Biol Chem. 1997 Apr 25;272(17):11495-502. doi: 10.1074/jbc.272.17.11495.
Five different monoclonal antibodies that immunoreact with RAP74, the large subunit of general transcription factor (TF) IIF, were produced and characterized. Using one of these antibodies, an affinity purification procedure was devised to isolate a human RNA polymerase II complex. This procedure is fast, simple, and reproducible and does not require extensive purification. The RNA polymerase II complex isolated using this procedure contains SRB (suppressor of RNA polymerase B) polypeptides, transcription factors IIE and IIF, limiting amounts of TFIIH, and the TATA-binding protein, but was devoid of TFIIB.
制备并鉴定了五种与通用转录因子（TF）IIF的大亚基RAP74发生免疫反应的不同单克隆抗体。使用其中一种抗体，设计了一种亲和纯化程序来分离人RNA聚合酶II复合物。该程序快速、简单且可重复，不需要广泛的纯化。使用该程序分离的RNA聚合酶II复合物包含SRB（RNA聚合酶B的抑制因子）多肽、转录因子IIE和IIF、有限量的TFIIH以及TATA结合蛋白，但不含TFIIB。
