Nucleotide sequence of the PaeR7 restriction/modification system and partial characterization of its protein products.
Nucleotide sequence of the PaeR7 restriction/modification system and partial characterization of its protein products.
复制标题
PaeR7 限制/修饰系统的核苷酸序列及其蛋白质产物的部分表征。
DOI：
10.1093/nar/13.23.8441
复制
发表时间：
1985
影响因子：
14.9
通讯作者：
T. Gingeras
中科院分区：
文献类型：
--
作者：
G. Thériault;P. H. Roy;K. A. Howard;J. Benner;J. Brooks;A. F. Waters;T. Gingeras
对铜绿假单胞菌PaeR7限制性修饰系统克隆的Bal31缺失实验表明，它作为一个操纵子排列，甲基化酶基因位于核酸内切酶基因之前。该操纵子的DNA序列与体外转录-翻译实验结果一致，分别预测了532个氨基酸（Mr = 59,260道尔顿）和246个氨基酸（Mr = 27,280道尔顿）与甲基化酶和核酸内切酶基因一致。这些预测值与纯化的、变性的PaeR7内切酶和甲基化酶蛋白的测量分子量一致。纯化的核酸内切酶氨基末端的前20个氨基酸与DNA序列预测的完全匹配。最后，基于几种PaeR7亚克隆的特性描述了噬菌体限制性表达的潜在调控机制。
Bal31 deletion experiments on clones of the PaeR7 restriction-modification system from Pseudomonas aeruginosa demonstrate that it is arranged as an operon, with the methylase gene preceding the endonuclease gene. The DNA sequence of this operon agrees with in vitro transcription-translation assays which predict proteins of 532 amino acids, Mr = 59,260 daltons, and 246 amino acids, Mr = 27,280 daltons, coincident with the methylase and endonuclease genes, respectively. These predicted values coincide with the measured molecular weights of the purified, denatured PaeR7 endonuclease and methylase proteins. The first twenty amino acids from the amino-terminus of the purified endonuclease exactly match those predicted from the DNA sequence. Finally, potential regulatory mechanisms for the expression of phage restriction are described based on the properties of several PaeR7 subclones.