Okamura S, Crane F, Messner H A, Mak T W
J Biol Chem. 1978 Jun 10;253(11):3765-7.
Terminal deoxynucleotidyltransferase is an enzyme which has been found to be associated with thymus cells, bone marrow cells, as well as leukocytes from patients with acute lymphoblastic leukemia and chronic myelocytic leukemia in blast crisis. We report here the purification of terminal deoxynucleotidyltransferase by an oligonucleotide affinity (oligo(dT)12-18 cellulose) column. By using a 35 to 70% (NH4)2SO4 cut, Sephacryl S200 column and an oligo(dT) cellulose column, terminal deoxynucleotidyltransferase has been purified from calf thymus cells to a specific activity of more than 8,500 units/mg of protein. The terminal deoxynucleotidyltransferase purified by this method contains no detectable DNA-dependent DNA polymerase or endonuclease activities. Furthermore, sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the enzyme appears to be homogeneous, with two polypeptides corresponding to the two subunits alpha (10,000) and beta (23,000) of terminal deoxynucleotidyltransferase. These data indicate that oligo(dT)12-18 cellulose can be used as a rapid and selective affinity column for the purification of terminal deoxynucleotidyltransferase.
末端脱氧核苷酸转移酶是一种已被发现与胸腺细胞、骨髓细胞以及急性淋巴细胞白血病和慢性粒细胞白血病急变期患者的白细胞相关的酶。我们在此报告通过寡核苷酸亲和（寡聚（dT）12 - 18纤维素）柱对末端脱氧核苷酸转移酶的纯化。通过使用35%至70%硫酸铵沉淀、Sephacryl S200柱和寡聚（dT）纤维素柱，已从小牛胸腺细胞中纯化出末端脱氧核苷酸转移酶，其比活性超过8500单位/毫克蛋白质。用这种方法纯化的末端脱氧核苷酸转移酶未检测到可检测的依赖DNA的DNA聚合酶或核酸内切酶活性。此外，十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳显示该酶似乎是均一的，有两条多肽链，分别对应末端脱氧核苷酸转移酶的两个亚基α（10000）和β（23000）。这些数据表明，寡聚（dT）12 - 18纤维素可用作纯化末端脱氧核苷酸转移酶的快速且选择性的亲和柱。
