Patent ID: 9580478
Date: 2017-02-28
CPC Classifications: A61K,A61P,C07K,C12N

Claim:
1. A method of making a recombinant mesophilic bacterium that is temperature-sensitive (TS), comprising: introducing into the genome of a mesophilic bacterium by homologous recombination a nucleic acid construct comprising a TS essential nucleic acid molecule from a psychrophilic bacterium flanked on both sides by a nucleic acid molecule homologous to a region of the mesophilic bacterium genome where the TS essential nucleic acid molecule from the psychrophilic bacterium will be inserted into the mesophilic bacterium genome, and functionally replacing the mesophilic bacterium's homolog of the TS essential nucleic acid molecule, thereby making a recombinant mesophilic bacterium that is TS, wherein a protein encoded by the TS essential nucleic acid molecule is operable at a temperature less than 30° C. and inoperable at a temperature greater than 30° C., and wherein the recombinant mesophilic bacterium that is TS has a restrictive temperature between 33° C. and 44° C.