Eniricerche S.p.a.

Antonio Mele

Guido Grandi

Salvatore Toma

Susanna Campagnoli

Marina Del Bue

Elisabetta Colletti

Renzo Nogarotto

pSM112 plasmid vector used for cloning heterologous proteins in Bacillus subtilis. A method of constructing plasmid pSM112 from plasmid pSM23 which contains the origin of replication of pUB110 and of pE194, said method comprising: a) plasmid pSM23 is linearized with XbaI restriction enzyme, b) the plasmid DNA is cyclized with T4 DNA ligase enzyme and plasmid pSM29 is isolated, c) the EcoRI-BamHI fragment of pSM29 containing the replication origin of pE194 is removed, d) the large EcoRI-BamHI fragment of pSM29 containing the replication origin of pUB110 is cyclized, and e) the plasmid pSM112 is isolated.

PSM112 plasmid vector for expression in bacillus subtilis

A description is given of a cloning vector useful for the expression and secretion of heterologous proteins in Bacillus subtilis, molecules of recombinant DNA formed by the said cloning vector, expediently cut at a restriction site located within the neutral protease gene, and by the sequence of heterologous DNA which codes for the protein of interest, Bacillus subtilis cells transformed with the said molecules and capable of expressing the sequence of heterologous DNA and of producing and secreting the protein coded by the said sequence of heterologous DNA in high yields. The proteins obtained in this manner are useful in the fields of pharmaceuticals and foods, or as compounds for the production of substances which are normally obtained by chemical synthesis.

Cloning vector, molecules of recombinant DNA, bacillus subtilis strains

Novel plasmid vectors are described, for expression in Escherichia coli and/or Bacillus subtilis, in which the gene which codes for a heterologous protein is placed under the control of a promoter of the erythromycin gene which permit the organisms transformed with the abovementioned plasmids to express the heterologous protein in high yields.

Growing community of inventors

Pavia, Italy

Antonio Mele