Abstract: Novel methods and novel industrial unicellular microorganism strains, particularly industrial Bacillus strains, are provided for enhanced production of endogenous and exogenous polypeptides. Cloning vehicles containing the gene expressing the polypeptide of interest are introduced into a compatible host. Transformed hosts harboring the introduced vehicle in a stable way by integration of the vehicle into the host cells chromosome are selected. Efficient transfer of the vehicle containing the gene of interest is achieved with the resulting industrial strain transformants being effective, stable producers of the desired polypeptide product.

Abstract: Novel methods and novel industrial unicellular microorganism strains, particularly industrial Bacillus strains, are provided for enhanced production of endogenous and exogenous polypeptides. Cloning vehicles containing the gene expressing the polypeptide of interest are introduced into a compatible host. Transformed hosts harboring the introduced vehicle in a stable way by integration of the vehicle into the host cells chromosome are selected Efficient transfer of the vehicle containing the gene of interest is achieved, with the resulting industrial strain transformants being effective, stable producers of the desired polypeptide product.

Abstract: Novel vectors are provided for identifying secretory signal sequences from DNA fragments of unicellular microorganisms. The plasmids comprise a multiple cloning site with restriction sites in reading frame with a structural gene which permits rapid screening of the secreted expression product. Optionally, the vectors may include a promoter region upstream from the multiple cloning site. The invention is exemplified with Bacillus. Specific secretory signal sequences have been isolated with those vectors, allowing for efficient secretion into the supernatant, and not just to the periplasmic space to provide proteins in economically high yields. Secretory sequences are provided superior to other previously known sequences.

Abstract: Novel vectors are provided for identifying secretory signal sequences from DNA fragments of unicellular microorganisms. The plasmids comprise a multiple cloning site with restriction sites in reading frame with a structural gene which permits rapid screening of the screted expression product. Optionally, the vectors may include a promoter region upstream from the multiple cloning site. The invention is exemplified with Bacillus. Specific secretory signal sequence have been isolated with those vectors, allowing for efficient secretion into the supernatant, and not just to the periplasmic space to provide proteins in economically high yields. Secretory sequences are provided superior to other previously known sequences.

Abstract: Novel constructs in plasmids are provided for evaluating the efficiency of expression and secretion of structural genes. The constructs provide for transcriptional and translational regulatory regions, a signal sequence and structural gene, which also may be readily excised and substituted, so as to allow for mixing and matching of regulatory regions, signal sequences and genes to evaluate regions for use in the expression of a desired peptide. Particularly, synthetic regions are provided which may be used with other synthetic regions or wild-type regions.Plasmids are provided for screening Bacillus genomic sequences for regulatory regions, particularly promoters employing a structural gene secreting an enzyme which can produce a product which allows for visual detection.