Construction and Use of a Broad-Host-Range Plasmid Expressing the lamB Gene for Utilization of Bacteriophage λ Vectors in the Marine Bacterium Vibrio harveyi

Abstract:

The remarkable success of Escherichia coli as a model organism in molecular genetics was dependent, among other things, on its susceptibility to genetic manipulation. Many versatile and sophisticated genetic tools for molecular biology studies are derived from bacteriophage λ. However, this bacteriophage is specific for E. coli, and thus λ-based techniques have been restricted to this bacterium. Plasmids expressing the E. coli gene coding for bacteriophage λ receptor were reported previously, and introduction of such plasmids into cells of some other bacteria made them sensitive to phage λ infection. However, we found that these systems were not efficient for Vibrio harveyi, one of the most frequently investigated species of marine bacteria. Here we describe construction of a broad-host-range plasmid expressing the lamB gene. Introduction of this plasmid to V. harveyi cells and expression of lamB made this strain susceptible to bacteriophage λ adsorption and λ DNA injection. Foreign genetic material could be introduced into cells of this strain using a cosmid vector.