Shizuya H, Richardson C C
Proc Natl Acad Sci U S A. 1974 May;71(5):1758-62. doi: 10.1073/pnas.71.5.1758.
We have developed a cell-free system to study bacteriophage lambda DNA replication. Maximal DNA synthesis in vitro requires the four deoxynucleoside triphosphates, ATP, and exogenous lambda DNA. DNA synthesis requires the products of the phage O and P genes but is not inhibited by lambda repressor. The kinetics of synthesis is linear for 10-15 min; however, the product of synthesis amounts to only 0.5-1% of the added template DNA. As judged by isopycnic analysis, extensive regions of the template are copied. Sedimentation analysis indicates that all of the product consists of short (11S) DNA chains. Fractions partially purified from lambdaO(+)P(+)-infected cell extracts will complement extracts prepared from lambdaO(-) or lambdaP(-)-infected cells.
我们开发了一种无细胞系统来研究噬菌体λDNA复制。体外最大DNA合成需要四种脱氧核苷三磷酸、ATP和外源λDNA。DNA合成需要噬菌体O和P基因的产物，但不受λ阻遏物抑制。合成动力学在10 - 15分钟内呈线性；然而，合成产物仅占添加模板DNA的0.5 - 1%。通过等密度分析判断，模板的大片段区域被复制。沉降分析表明，所有产物均由短（11S）DNA链组成。从λO(+)P(+)感染细胞提取物中部分纯化的组分将补充从λO(-)或λP(-)感染细胞制备的提取物。
