Graham S, Yoneda Y, Young F E
Gene. 1979 Sep;7(1):69-77. doi: 10.1016/0378-1119(79)90043-x.
Spontaneous deletion mutants of the bacteriophage SPO2, which are viable and retain their temperate character, were isolated using a heat-EDTA enrichment step. They were identified by endonuclease digestion and agarose-gel electrophoresis of phage DNA. Two of the nine mutants were characterized in detail. Both mutants have a 2.3 Md deletion removing the single BglII site and two of the XbaI fragments. The deletion extends 1.0 Md to one side of the former BglII site and 1.3 Md on the other side. This region of the SPO2 genome is non-essential for either lysogeny or viable phage production and thus is a suitable region for the insertion of exogenous DNA fragments.
利用热-乙二胺四乙酸(EDTA)富集步骤,分离出了噬菌体SPO2的自发缺失突变体,这些突变体具有活性并保留其温和特性。通过噬菌体DNA的核酸内切酶消化和琼脂糖凝胶电泳对它们进行鉴定。对九个突变体中的两个进行了详细表征。两个突变体都有一个2.3兆达尔(Md)的缺失,去除了单个BglII位点和两个XbaI片段。该缺失在原来BglII位点的一侧延伸1.0 Md,在另一侧延伸1.3 Md。SPO2基因组的这个区域对于溶原性或有活性噬菌体的产生都不是必需的,因此是插入外源DNA片段的合适区域。