Method for stabilizing recombinant DNA molecules and enhancing the expression thereof in strains of Bacillus
AssigneesAgency of Industrial Science and Technology, Ministry of International Trade & Industry
InventorsYoshio Furutani, Masaru Honjo, Kazuaki Manabe, Hiroaki Shimada, Noboru Tomioka
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Abstract
A recombinant DNA molecule, a portion of which includes a vector whose replication is initiated independently of the temperature-sensitive mutant gene of a temperature-sensitive mutant strain of the genus Bacillus as host is introduced into the host. The host is then cultured at a temperature which does not completely inhibit chromosomal DNA replication. By this process, the recombinant DNA molecule can be stabilized in the host and its expression can be enhanced.
Claims17
A method for stabilizing and enhancing expression of recombinant DNA molecules comprising
- (a) providing a temperature-sensitive mutant strain of genus Bacillus in which an initiation of chromosomal DNA replication is inhibited in a specific temperature range,
- (b) introducing a recombinant DNA molecule into the temperature-sensitive mutant strain having a vector with a replication origin of pUB110 in which replication is initiated independently of a temperature-sensitive mutant gene of the temperature-sensitive mutant strain and
- (c) culturing the temperature-sensitive mutant strain in a temperature range from and including 30.degree. C. to below 45.degree. C. and where initiation of chromosomal DNA replication of the temperature-sensitive mutant strain is partially inhibited,
- such that said recombinant DNA molecules during said chromosomal DNA replication are stably retained and a copy number of said recombinant DNA molecules is increased.
Dependencies are inferred from the claim text.