Watabe K, Ito J
Nucleic Acids Res. 1983 Dec 10;11(23):8333-42. doi: 10.1093/nar/11.23.8333.
A novel DNA polymerase induced by Bacillus subtilis bacteriophage phi 29 has been identified. This polymerase can be separated from host DNA polymerase, by fractionation of extracts prepared from phage infected cells, using phosphocellulose chromatography. The isolated polymerase prefers poly(dA)oligo(dT) as template. The DNA polymerase isolated from the cells infected with a gene 2 temperature sensitive mutant (ts2) showed greater heat-lability than that induced by wild type phi 29. The ts2 DNA polymerase was also thermolabile for its activity in the formation of a covalent complex between phi 29 terminal protein and dAMP, the initiation step of phi 29 DNA replication. These findings indicate that gene 2 is the structural gene for a phi 29 DNA polymerase required for the complex formation step of DNA initiation.
已鉴定出一种由枯草芽孢杆菌噬菌体φ29诱导产生的新型DNA聚合酶。通过用磷酸纤维素色谱法对噬菌体感染细胞制备的提取物进行分级分离，可将这种聚合酶与宿主DNA聚合酶分离。分离出的聚合酶更倾向于以聚（dA）寡聚（dT）作为模板。从感染了基因2温度敏感突变体（ts2）的细胞中分离出的DNA聚合酶比野生型φ29诱导产生的DNA聚合酶表现出更高的热不稳定性。ts2 DNA聚合酶在φ29末端蛋白与dAMP形成共价复合物（即φ29 DNA复制的起始步骤）的过程中，其活性也表现出热不稳定性。这些发现表明，基因2是DNA起始复合物形成步骤所需的φ29 DNA聚合酶的结构基因。
