Stable gene amplification in prokaryotic chromosomal dna

C - Chemistry – Metallurgy – 12 – N

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C12N 15/90 (2006.01) C12N 1/21 (2006.01) C12N 9/28 (2006.01) C12N 9/54 (2006.01) C12N 15/67 (2006.01) C12N 15/75 (2006.01)

Patent

CA 1327175

ABSTRACT Transformed prokaryotic hosts are provided compris- ing two or more copies of a DNA sequence stably maintained in their chromosome, said sequence comprising a gene encoding a polypeptide of interest, wherein said copies are separated by endogenous chromosomal DNA sequences. Methods are also pro- vided for producing said transformed host strains. Said transformed host strains are capable of increased production of the polypeptide of interest compared to host strains which already produce said polypeptide. Preferred host strains are Bacillus novo species PB92 which produces a high-alkaline pro- teolytic enzyme and Bacillus licheniformis T5 which produce a thermostable alpha-amylase, and mutants and variants of said strains. Preferred polypeptide encoding genes are the pro- tease encoding gene originating from Bacillus PB92 and the alpha-amylase encoding gene originating from Bacillus lichen- iformis strain T5.

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