Miyashita S, Hirochika H, Ikeda J E, Hashiba T
Laboratory of Plant Pathology, Faculty of Agriculture, Tohoku University, Sendai, Japan.
Mol Gen Genet. 1990 Jan;220(2):165-71. doi: 10.1007/BF00260476.
Three linear DNA plasmids were found in isolate RI-64 of anastomosis group 4 (AG-4) of Rhizoctonia solani. These plasmids, designated pRS64-1, -2, and -3, possessed the same size of 2.7 kb. Restriction mapping and Southern hybridization analysis of pRS64-1, -2, and -3 revealed the presence of homologous regions at both termini. The plasmid DNAs were resistant to both 3'-exonuclease and 5'-exonuclease even after treatment with proteinase K or alkali. The length of both terminal fragments that were generated by restriction endonuclease digestion was doubled under the denaturation condition, indicating that the linear plasmid DNAs have hairpin loops at both termini. Southern blotting analysis of total DNA showed the presence of two types of dimeric forms of pRS64 DNA. One is a head-to-head dimer and the other is a tail-to-tail dimer. The role of these unique DNA structures in replication of the plasmids is discussed.
在立枯丝核菌融合群4(AG - 4)的分离株RI - 64中发现了三种线性DNA质粒。这些质粒被命名为pRS64 - 1、- 2和- 3,大小均为2.7 kb。对pRS64 - 1、- 2和- 3进行限制性图谱分析和Southern杂交分析,结果显示其两端均存在同源区域。即使在用蛋白酶K或碱处理后,质粒DNA对3' - 外切核酸酶和5' - 外切核酸酶均具有抗性。在变性条件下,经限制性内切酶消化产生的两个末端片段的长度加倍,这表明线性质粒DNA两端具有发夹环。对总DNA进行的Southern印迹分析显示存在两种类型的pRS64 DNA二聚体形式。一种是头对头二聚体,另一种是尾对尾二聚体。本文讨论了这些独特DNA结构在质粒复制中的作用。