Identification of genes in pseudomonas bacteria

C - Chemistry – Metallurgy – 07 – H

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195/1.18, 195/1.

C07H 21/04 (2006.01) C12N 1/20 (2006.01) C12N 15/78 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 1264685

ABSTRACT OF THE DISCLOSURE Genetic analysis and modification of Pseudomonas bacteria is accomplished using a suicide plasmid which includes genes for replication in a narrow range of host cells and genes for conjugal transfer of the plasmid into a broad range of bacterial hosts, including Pseudomonas. The genetic donor is E coli and the plasmid carries a transposon having at least one antibiotic resistance marker, e.g. kanamycin resistance or tetracycline resistance, conferring thereon an antibiotic resistance not found in the Pseudomonas host or in the remainder of the suicide plasmid. The suicide vector is constructed and introduced into E coli cells, in which it replicates. Then it is introduced into Pseudomonas cells by conjugal transfer, whereupon the transposon inserts into the DNA of the recipient, deactivating or altering the activity of a gene therein, whilst the residue of the vector is eliminated and does not replicate. By this means, the operative symbiotic genes in the Pseudomonas may be identified, by comparison of the genetic behaviour of the modified Pseudomonas cells so formed with wild type Pseudomonas cells.

469745

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