Takada-Takayama R, Suzuki M, Enomoto T, Hanaoka F, Ui M
Department of Physiological Chemistry, Faculty of Pharmaceutical Sciences, University of Tokyo, Japan.
FEBS Lett. 1990 Oct 29;273(1-2):27-30. doi: 10.1016/0014-5793(90)81043-n.
A simple method was developed for the isolation of primase-free DNA polymerase-alpha from the DNA polymerase-alpha-primase complex of mouse FM3A cells. The polymerase was separated from primase subunits by chromatography on a single-stranded DNA-cellulose column in the presence of 50% etylene glycol. The primase-free DNA polymerase-alpha contained two polypeptides with molecular masses of 180,000 and 68,000. Analysis of the DNA products with poly(dA)-oligo(dT)10 as template-primer revealed that both primase-free DNA polymerase-alpha and the DNA polymerase-alpha-primase complex predominantly synthesized short DNA with less than 30 nucleotides, but that the DNA polymerase-alpha-primase complex also synthesized some longer DNA with more than 300-400 nucleotides.
已开发出一种从小鼠FM3A细胞的DNA聚合酶α-引发酶复合物中分离无引发酶的DNA聚合酶α的简单方法。在50%乙二醇存在下，通过单链DNA-纤维素柱层析将聚合酶与引发酶亚基分离。无引发酶的DNA聚合酶α包含两条分子量分别为180,000和68,000的多肽。以聚（dA）-寡聚（dT）10为模板-引物对DNA产物进行分析表明，无引发酶的DNA聚合酶α和DNA聚合酶α-引发酶复合物主要合成少于30个核苷酸的短DNA，但DNA聚合酶α-引发酶复合物也合成一些超过300-400个核苷酸的较长DNA。
