Complementation assay of primer protein: gene expression systems of plasmid vectors support the infection of suppressor sensitive mutant phages M2 and phi 29.

Idengaku zasshi Pub Date : 1993-08-01 DOI:10.1266/jjg.68.243
T Kishi, K Miura, K Matsumoto, H Hirokawa
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引用次数: 5

Abstract

Two different expression systems of genes of primer proteins (pE for phage M2, and p3 for phi 29) were constructed to study the protein primed DNA replication of Bacillus phages M2 and phi 29. In one system, expression of the genes was under the control of the inducible spac promoter, whereas in the other system, the expression was under the control of the constitutive promoter in plasmid pUB110. Complementation tests in vivo were performed between the primer proteins expressed by these systems and mutant phages having suppressor sensitive mutations in the genes of the primer proteins. The phages M2 susE and phi 29 sus3 were complemented by pE and p3 expressed by the systems, respectively. However, the complementation and apparent phage DNA synthesis were not detected in the combinations between susE of phage M2 and p3 of phage phi 29, and vice versa. Although pE and p3 proteins exhibited structurally and functionally similar characteristics, these proteins showed species specificity in the protein primed DNA replication of bacteriophages M2 and phi 29.

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引物蛋白的互补实验:质粒载体的基因表达系统支持抑制敏感突变体M2和phi 29的感染。
构建了噬菌体M2的引物蛋白基因pE和phi 29的引物蛋白p3两个不同的表达体系,研究了噬菌体M2和phi 29的蛋白引物DNA复制。在一个系统中,基因的表达受诱导空间启动子的控制,而在另一个系统中,基因的表达受质粒pUB110中的组成启动子的控制。在体内进行了由这些系统表达的引物蛋白和在引物蛋白基因中具有抑制敏感突变的突变噬菌体之间的互补试验。噬菌体M2 susE和phi 29 sus3分别被系统表达的pE和p3补充。而噬菌体M2的susE与噬菌体phi 29的p3的组合没有发现互补和明显的噬菌体DNA合成,反之亦然。尽管pE和p3蛋白在结构和功能上具有相似的特征,但它们在噬菌体M2和phi 29的蛋白引物DNA复制中表现出物种特异性。
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