Method for stabilizing recombinant DNA molecules and enhancing t

Chemistry: molecular biology and microbiology – Treatment of micro-organisms or enzymes with electrical or... – Modification of viruses

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935 43, 935 74, 435 68, C12N 1500, C12P 2100

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047313277

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.

REFERENCES:
patent: 4495287 (1985-01-01), Uhlin et al.
Shivakumar, et al., Plasmid 1:405-416 (1978).
Winston, et al., "DNA-Membrane Association is Necessary for Initiation of Chromosomal and Plasmid Replication in Bacillus subfilis" Proc. Natl. Acad Sci vol. 77(5), pp. 2834-2838, May 1980.
Principles of Gene Manipulation and Introduction to Genetic Engineering, 1980 Blackwell Scientific Publications, pp. 48 through 59.
Construction and Properties of Chimeric Plasmids in Bacillus subtilis, Gryczan et al., 1977, pp. 1423 through 1432.
Segregational Instability of pUB110-Derived Recombinant Plasmids in Bacillus subtilis, Bron et al., 1985, pp. 235 through 244.
The Third International Conference on Genetics and Biotechnology of Bacilli, 1985, A. T. Ganesan (Stanford), pp. 3, 4, and 51.
Ehrlich, S. D. et al., "DNA Recombination in Plasmids and the Chromosome of Bacillus subtilis," A. T. Ganesan & J. A. Hoch (Eds), Bacillus Molecular Genetics and Biotechnology Applications, Academic Press, Inc., Orlando, (1986), pp. 27-28.
Hardy, K. G., "Bacillus Cloning Methods", D. M. Glover (Ed.), DNA Cloning Volume II a Practical Approach, Irl Press, Oxford, pp. 1-5.

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