de Vries G E, Raymond C K, Ludwig R A
Proc Natl Acad Sci U S A. 1984 Oct;81(19):6080-4. doi: 10.1073/pnas.81.19.6080.
A set of plasmids has been constructed that carry a constitutive lamB gene (LamBc phenotype) from Escherichia coli and that confer functional phage lambda receptors to bacteria other than E. coli. This E. coli LamBc strain has been selected to escape both maltose-inducible and glucose-repressible control. Constitutivity results from an IS-3 insertion, carrying a mobile promoter, proximal to lamB. The LamBc DNA has been cloned into both broad and narrow host-range plasmids, and the resulting pTROY plasmids have been transferred to diverse bacteria. Both Salmonella typhimurium/pTROY and Klebsiella pneumoniae/pTROY strains efficiently adsorb phage lambda; Pseudomonas aeruginosa/pTROY strains do not. Introduction of a functional E. coli LamB protein into foreign bacterial will allow these bacteria carrying pTROY plasmids to be infected by phage lambda recombinant DNA libraries, phage lambda::Tn insertion mutagenesis vectors, and in vivo lambda-packaged cosmids.
已经构建了一组质粒,这些质粒携带来自大肠杆菌的组成型lamB基因(LamBc表型),并赋予除大肠杆菌以外的细菌功能性噬菌体λ受体。选择这种大肠杆菌LamBc菌株以逃避麦芽糖诱导型和葡萄糖抑制型控制。组成性是由位于lamB近端的携带移动启动子的IS-3插入导致的。LamBc DNA已被克隆到广泛宿主范围和窄宿主范围的质粒中,所得的pTROY质粒已被转移到多种细菌中。鼠伤寒沙门氏菌/pTROY和肺炎克雷伯菌/pTROY菌株都能有效吸附噬菌体λ;铜绿假单胞菌/pTROY菌株则不能。将功能性大肠杆菌LamB蛋白引入外来细菌将使携带pTROY质粒的这些细菌能够被噬菌体λ重组DNA文库、噬菌体λ::Tn插入诱变载体以及体内λ包装的黏粒感染。