Valio Ltd.

Oy Alko Ab

Genesit Oy

Valio, Finnish Co-Operative Dairies' Association

The Finnish National Public Health Institute

Ilkka Palva

Mervi Sibakov

Willem Meindert De Vos

Soile Tynkkynen

Atte Von Wright

Ingemar Von Ossowski

Tuomas Salusjärvi

Airi Palva

Reetta Satokari

Satu Vesterlund

Teija Koivula

Justus Reunanen

Matti Kankainen

The present invention relates to the fields of life sciences and food, feed or pharmaceutical industry. Specifically, the invention relates to novel peptides, pilus structures, polynucleotides as well as vectors, host cells, products and pharmaceutical compositions comprising the polynucleotides, peptides or pilus structures. The invention also relates to gene clusters and antibodies. Furthermore, the present invention relates to methods for producing the peptides or pilus structures or producing the products comprising the peptides or pilus structures. Furthermore, the present invention relates to treatments as well as uses and methods for screening bacterial strains, for reducing or inhibiting the adhesion of pathogenic bacteria, promoting the adhesion of bacterial cells to the mucus and for modifying immune response in a subject. Still, the present invention relates to methods for detecting probiotic bacterial strains or pathogen strains.

Peptides and methods for producing them

DNA sequences, derived from Lactococcus lactis subsp. lactis, are useful as promoters and promoter/secretion promoting signals for heterologous or homologous expression in Gram-positive bacteria. In another aspect, the invention relates to vectors, such as plasmids, comprising the sequences of the present invention, and to host cells transformed with such vectors. Yet additional aspects of the present invention are related to methods for producing desired heterologous or homologous peptides or proteins employing the sequences, vectors, or transformed hosts of the invention. By means of the invention, greatly improved heterologous and homologous expression and secretion may be achieved in E. coli and in Gram-positive bacteria such as B. subtilis, Lactococcus and Lactobacillus.

Lactococcus promoters and signal sequences for heterologous gene

The invention relates to recombinant DNA molecules and to methods for producing proteins by means of said molecules. Particularly, the present invention relates to recombinant DNA molecules which are capable of being synthesized in Bacillus strain bacteria comprising the regulation and deleted non-functional signal sequence of the a-amylase gene of B. amyloliquefaciens, or a substantial part thereof, to which sequence a structural gene of any desired homologous or heterologous protein or peptide may be joined. These recombinant DNA molecules can be used, for example, to achieve intracellular expression of any desired protein or peptide in Bacillus strain bacteria.

Expression vectors and methods for intracellular protein production in

DNA sequences, derived from Lactococcus lactis subsp. lactis, are useful as promoters nd promoter/secretion promoting signals for heterologous or homologous expression in Gram-positive bacteria. In another aspect, the invention relates to vectors, such as plasmids, comprising the sequences of the present invention, and to host cells transformed with such vectors. Yet additional aspects of the present invention are related to methods for producing desired heterologous or homologous peptides or proteins employing the sequences, vectors, or transformed hosts of the invention. By means of the invention, greatly improved heterologous and homologous expression and secretion may be achieved in E. coli and in Gram-positive bacteria such as B. subtilis, Lactococci and Lactobacillus.

Lactococcus promoter and signal sequences for expression in bacteria

A selected protein or protein part is prepared in Bacillus strain bacteria by joining DNA encoding the selected protein or protein part essential to its biological activity to a bacterium gene. The Bacillus amyloliquefaciens gene for .alpha.-amylase is cleaved at a location after the excretion signal following the regulation sequence, or at a location after a part essential with respect to its excretion. The cleaved gene is joined to a plasmid present in Bacillus strain bacteria in several copies, and the DNA sequence encoding the selected protein or protein part essential to its biological activity is joined to the cleavage site by recombinant DNA techniques. Bacillus strain host bacteria are transformed with the recombinant DNA molecules so obtained, and the transformed host bacteria cultivated to produce the selected protein or protein part.

Method for the preparation of a selected protein or a part thereof in

The invention relates to recombinant DNA molecules and to methods for producing proteins by means of said molecules. Particularly, the present invention relates to recombinant DNA molecules which are synthetized in Bacillus strain bacteria and are characterized by DNA which codes for exoenzymes excreted by a bacterium of the Bacillus strain and which are present in tens of copies in Bacillus strain bacteria, as well as to recombinant DNA molecules which are modified from the above recombinant DNA molecules and are characterized by DNA containing the regulation and excretion signals of the .alpha.-amylase gene of B. amyloliquefaciens, to which signals a gene of any protein can be joined. These recombinant DNA molecules can be used, for example, to improve the production of .alpha.-amylase in Bacillus strain bacteria, and their modifications to produce any protein in Bacillus strain bacteria.

Recombinant DNA-molecules and method for protein production

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Helsinki, Finland

Ilkka Palva