Abstract: The present invention provides an anti-VEGF human monoclonal antibody capable of binding to human VEGF to neutralize its activity, a cell strain producing said human monoclonal antibody, and a vascularization inhibitor containing said human monoclonal antibody. Hybridoma strains VA01 and BL2 producing the anti-VEGF human monoclonal antibody can also be provided according to the present invention by transformation with Epstein-Barr Virus (EBV) of human lymphocyte producing anti-human VEGF antibody and then fusing the transformant cells with human myeloma cells. The monoclonal antibody of the present invention can serve as a vascularization inhibitor which is useful for human because it is derived from human and capable of binding specifically to VEGF participating in vascularization, to inhibit the migration or proliferation of vascular endothelial cells.

Abstract: A neutral protease gene of Bacillus amyloliquefaciens is cloned, which gene comprises the promoter region, the ribosome binding region, the region involved in the secretion of the neutral protease, the region consisting of the structural gene for the neutral protease, and the terminator region. Each of the regions is useful as a material for construction of a recombinant DNA used for the production of proteins by culturing a host bacterium transformed with the recombinant DNA. For example, the extracellular production of neutral protease in a large amount can be accomplished by culturing B. subtilis transformed with a recombinant DNA comprising pUB110 and the neutral protease gene.

Abstract: Residual extracellular protease activities of a Bacillus subtilis strain can be further reduced by introducing a gene for the stimulation of extracellular protease levels into the genomic DNA of the strain.The resultant strain whose extracellular protease activities are further reduced is transformed with a recombinant plasmid for secretion of a desired protein and the desired protein can be efficiently produced and accumulated in the culture medium in a large amount. The accumulated desired protein can be easily recovered from the culture medium.For example, the accumulation level of human growth hormone secreted by a B. subtilis MT-430 strain carrying recombinant DNA phGH427 for the expression and secretion of human growth hormone can be increased five to tenfold as a result of the above treatment for reduction of the residual extracellular protease activities of the extracellular protease-activity-deficient strain.

Abstract: A expression-secretion vector is constructed by using a DNA sequence comprising a DNA fragment obtained by subjecting a DNA sequence consisting of a region involved in the expression of a gene coding for an extracellular enzyme of a bacterium of the genus Bacillus and a region involved in the secretion of the enzyme thus expressed to a deletion in the region involved in the secretion which can enhance the extracellular production of a protein dependent on the DNA fragment. By transforming a host bacterium with a recombinant DNA molecule formed by inserting a DNA fragment comprising a gene coding for a desired protein into the expression-secretion vector and then culturing the transformed host, the extracellular production of the desired protein in a large amount can be accomplished.