Dna Plant Technology Corporation

William T Tucker

Neal I Gutterson

Paul K Wolber

Viable bacteria may be detected in biological samples by exposing bacterial cultures obtained from the samples to transducing particles having a known host range. Such transducing particles carry a heterologous gene capable of altering the phenotype of the bacteria in a readily detectable manner. For example, the transducing particles may carry an ice nucleation gene and the alteration of phenotype may be detected using an ice nucleation assay. By employing a panel of phage, unknown bacteria nmay be typed based on the pattern of reactivity observed. The transducing particles may be prepared by introducing a synthetic transposable element carrying the heterologous gene to a host carrying a prophage having the desired host range. After transposition, the host may be induced to a lytic cycle to release the transducing particles carrying the heterologous gene.

Transducing particles and methods for their production

This invention relates to a new method for inserting heterologous genes into the genome of a bacteria using a combined plasmid. The combined plasmid provides a cis complementation of transposase genes and transposable elements. The method involves the homologous recombination of a carrier plasmid and a functions plasmid to form the combined plasmid. The carrier plasmid contains a transposable element which flanks a generic expression cassette. The functions plasmid comprises transposase genes which complement the transposable element on the carrier plasmid. The combined plasmid is then transferred to a recipient and the recipient is monitored for integration of the generic expression cassette into the genome. The combined plasmid is preferably created by an in vivo homologous recombination of the carrier and functions plasmids.

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Oakland, CA, United States of America

William T Tucker