单链核酸内切酶从膜复合物中释放大肠杆菌DNA。
Release of Escherichia coli DNA from membrane complexes by single-strand endonucleases.
作者信息
Abe M, Brown C, Hendrickson W G, Boyd D H, Clifford P, Cote R H, Schaechter M
出版信息
Proc Natl Acad Sci U S A. 1977 Jul;74(7):2756-60. doi: 10.1073/pnas.74.7.2756.
Abstract
Treatment of gently prepared lysates of Escherichia coli with single-strand-specific endonuclease (SI or from mung beans) results in the release of about 90% of the DNA from membranes, as determined by the M band technique. The released DNA has an average molecular weight of about 1.2 X 10(8). Data obtained with endonuclease S1 fit a mathematical model in which substrate sites are at or near membrane attachment sites. Data obtained with pancreatic deoxyribonuclease or x-rays fit a model for double-strand breaks at random sites along the DNA. Fitting data to these models, we estimate that there are 18+/-5 membrane attachment sites. The DNA remaining after S1 nuclease treatment is enriched for the region near the origin of chromosome replication. Therefore, attachment at this region near the origin of chromosome replication. Therefore, attachment at this region appears to be chemically different from that at the other sites along the DNA.
摘要
用单链特异性核酸内切酶（S1或绿豆核酸酶）处理经温和制备的大肠杆菌裂解物，通过M带技术测定，约90%的DNA从膜上释放出来。释放出的DNA平均分子量约为1.2×10⁸ 。用核酸酶S1获得的数据符合一个数学模型，其中底物位点位于膜附着位点处或其附近。用胰腺脱氧核糖核酸酶或X射线获得的数据符合一个关于DNA上随机位点双链断裂的模型。将数据拟合到这些模型中，我们估计有18±5个膜附着位点。S1核酸酶处理后剩余的DNA在染色体复制起点附近的区域富集。因此，在染色体复制起点附近的这个区域的附着似乎在化学性质上与DNA上其他位点的附着不同。
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