Patent ID: 6066473
Filing Date: 2000-05-23
Classification: C12N

Abstract:
A method of introducing a DNA construct encoding a polypeptide of interest into a cell of a Bacillus sp., comprising mixing (a) a population of bacterial donor cells harboring (i) a plasmid comprising the DNA construct and at least one cis-acting DNA sequence required for the transfer of said plasmid by conjugation in the presence of a trans-acting mobilizing element, and (ii) at least one DNA sequence encoding said trans-acting mobilizing element, and (b) a population of Bacillus sp. recipient cells, under conditions allowing the plasmid to be transferred from the population of donor cells to the population of recipient cells by conjugation, wherein the plasmid, following transfer into the recipient cell, is integrated into the genome of the recipient cell and the DNA construct encoding the polypeptide of interest is introduced into the recipient Bacillus sp. cell, and wherein the recipient cell is an alkalophilic Bacillus sp., an industrial Bacillus strain and/or a mutant Bacillus strain having been mutated so that the polypeptide of interest is produced and secreted in an increased yield as compared to the corresponding unmutated strain.