Guilfoyle T J, Hagen G, Malcolm S
J Biol Chem. 1984 Jan 10;259(1):640-8.
Antibodies were raised against native soybean RNA polymerase II and individual subunits of RNA polymerase II purified from soybean, cauliflower, and wheat. These antibodies were used to study the immunological relationships of plant RNA polymerases I, II, and III at the subunit level. RNA polymerases I and II from soybean, I, II, and III from cauliflower and wheat, and II from turnip were purified to homogeneity, and enzyme subunits were separated on polyacrylamide gels containing sodium dodecyl sulfate. Separated RNA polymerase subunits were electrophoretically transferred to nitrocellulose, reacted with antibodies, and the immunoglobulin complexes were revealed by reaction with 125I-labeled Protein A. Antibodies directed against native soybean RNA polymerase II bind to all or most of the subunits of soybean and wheat RNA polymerase II and to the putative common subunits in RNA polymerases I and III. Antibodies directed against individual subunits of RNA polymerase II enzymes react specifically with the subunit to which the antibody was raised and to related or identical subunits in RNA polymerases I and III. These studies confirm the presence of common subunits in the three classes of nuclear RNA polymerase and provide information on analogous or related subunits within each class of enzyme purified from different plant species.
制备了针对天然大豆RNA聚合酶II以及从大豆、花椰菜和小麦中纯化得到的RNA聚合酶II各个亚基的抗体。这些抗体被用于在亚基水平上研究植物RNA聚合酶I、II和III的免疫关系。将大豆的RNA聚合酶I和II、花椰菜和小麦的RNA聚合酶I、II和III以及芜菁的RNA聚合酶II纯化至均一状态，并在含有十二烷基硫酸钠的聚丙烯酰胺凝胶上分离酶亚基。将分离得到的RNA聚合酶亚基通过电泳转移至硝酸纤维素膜上，与抗体反应，然后通过与125I标记的蛋白A反应来显示免疫球蛋白复合物。针对天然大豆RNA聚合酶II的抗体与大豆和小麦RNA聚合酶II的所有或大部分亚基以及RNA聚合酶I和III中假定的共同亚基结合。针对RNA聚合酶II酶各个亚基的抗体与产生该抗体的亚基以及RNA聚合酶I和III中相关或相同的亚基发生特异性反应。这些研究证实了三类核RNA聚合酶中存在共同亚基，并提供了从不同植物物种纯化得到的每类酶中类似或相关亚基的信息。
