Gene tandem-mediated selection of coliphage λ-receptive Agrobacterium, Pseudomonas , and Rhizobium strains

Abstract

Agrobacterium, Pseudomonas, and Rhizobium spp. have been made receptive to coliphage lambda. To achieve this, recombinant (pTROY) plasmids carrying a constitutive Escherichia coli lamB gene encoding the lambda receptor and expressed from an insertion sequence 3 (IS3) promoter were introduced into various bacteria. Because the wild-type lambda receptor was not expressed in these bacteria, a procedure called the lamB gene tandem protocol was used to select lamB alleles that expressed the lambda receptor. This gene tandem protocol may have general use in adapting genes with nonselectable traits to different organisms. Agrobacterium, Pseudomonas, and Rhizobium strains carrying pTROY41613, which encoded a gene tandem-selected lambda receptor, could be quantitatively transduced with lambda-packaged cosmids. With the ability to confer lambda receptivity on organisms, phage lambda now becomes a general DNA delivery agent.