Csonka L N, Howe M M, Ingraham J L, Pierson L S, Turnbough C L
J Bacteriol. 1981 Jan;145(1):299-305. doi: 10.1128/jb.145.1.299-305.1981.
A procedure was developed for introducing the coliphage Mu d1 (Apr lac) into Salmonella typhimurium in order to construct gene fusions that place the structural genes of the lac operon under the control of the promoter-regulatory region of other genes. To introduce Mu d1 from Escherichia coli K-12 into S. typhimurium, which is normally not a host for Mu, we first constructed an E. coli double lysogen carrying the defective Mu d1 phage and a Mu-P1 hybrid helper phage (MuhP1) that confers the P1 host range. A lysate prepared from this strain was used to infect a P1-sensitive (i.e., galE), restriction-deficient, modification-proficient strain of S. typhimurium, and a double lysogen carrying Mu d1 and MuhP1 was isolated. Induction of the latter strain produced lysates capable of infecting and generating gene fusions in P1-sensitive strains of S. typhimurium. In this paper we describe the construction of pyr::lac fusions by this technique.
为了构建基因融合体,使乳糖操纵子的结构基因受其他基因的启动子调控区控制,我们开发了一种将大肠杆菌噬菌体Mu d1(Apr lac)导入鼠伤寒沙门氏菌的方法。为了将Mu d1从大肠杆菌K-12导入通常不是Mu宿主的鼠伤寒沙门氏菌,我们首先构建了一个携带缺陷型Mu d1噬菌体和赋予P1宿主范围的Mu-P1杂交辅助噬菌体(MuhP1)的大肠杆菌双溶原菌。从该菌株制备的裂解物用于感染一株对P1敏感(即galE)、限制缺陷、修饰 proficient的鼠伤寒沙门氏菌菌株,并分离出携带Mu d1和MuhP1的双溶原菌。诱导后一种菌株产生的裂解物能够感染鼠伤寒沙门氏菌的P1敏感菌株并产生基因融合体。在本文中,我们描述了通过这种技术构建pyr::lac融合体的过程。