Process for amplifying the expression of a specific gene in baci

Details Process for amplifying the expression of a specific gene in baci Process for amplifying the expression of a specific gene in baci

1987-12-18

1990-09-25

Martinell, James

Chemistry: molecular biology and microbiology

Treatment of micro-organisms or enzymes with electrical or...

Modification of viruses

435 691, 435 712, 43525231, 435832, 435839, 935 42, 935 56, 935 74, C12N 1500, C12N 100, C12N 120, C12P 2100

Patent

active

049593161

ABSTRACT:
The present invention relates to a process for preparing a strain of Bacillus, in the chromosome of which a specific gene has been amplified, in which process: (a) at least one plasmid integration vector bearing the said gene is integrated in the Bacillus chromosome so as to create in the chromosome: at least one DNA sequence, known as an amplifiable unit, which contains at least the said gene and its expression elements and, at each end, two sequences which are identical in the direct sense; the amplifiable unit which codes, furthermore, for a selectable gene; (b) the strains of Bacillus obtained are then selected by culture on a selection medium corresponding to the selectable gene, and the strains are withdrawn which have the phenotype corresponding to the presence of an increased number of copies of the said gene relative to the bacterial population before selection.

REFERENCES:
patent: 4336336 (1982-06-01), Silhavy et al.
Buchanan et al., (ed.), Bergey's Manual of Determinative Bacteriology, Eighth Edition, 1974, Williams & Wilkins Co., Baltimore, pp. 529-533.
Young, "Gene Amplification in Bacillus Subtilis: The Establishment of Multiple Tandemly-Repeated Copies of Heterologous DNA Segment in the Bacterial Chromosome", Chem. Abstr. 102: 198792a, (1985) of Genet. Biotechnol. Bacilli, Ganesan et al., (ed.), (1984).
Bonamy et al., "Cloning and Expression of Bacillus Subtilis Spore Genes", Molec. Gen. Genet. 188: 202 (1982).
Chem. Abstr. 102: 1392r (1985) of JP 59,151,889 (1984).
Saunders et al., "Use of Chromosomal Integration in the Establishment and Expression of blaZ, a Staphylococcus Aurea Beta-Lactamase Gene in Bacillus Subtilis", J. Bacteriol. 157: 718 (1984).
Duncan et al., "Mechanism for Integrating Foreign DNA during Transformation of Bacillus Subtilis", Proc. Natl. Acad. Sci., U.S.A., 75: 3664 (1978).
Galizzi et al., "Integration and Excision of a Plasmid in Bacillus Subtilis", Mol. Gen. Genet. 182: 99 (1981).
Williams et al., "Expression of E. Coli trp Genes Cloned in B. Subtilis", in Molecular Cloning and Gene Regulation in Bacilli, 1982, Ganesan et al., (ed.), Academic Press, New York, pp. 91-96.
Gaillard, Excision Sequences in the Mitochondrial Genome of Yeast, Nature, vol. 283, Jan., 1980, p. 218.

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