| 摘要 |
PURPOSE:To secrete and produce a protein by genetic engineering means in high efficiency by reacting a specific sequence originated from a heat-resistant neutral protease gene of bacterium of genus Bacillus in a cell of Bacillus, etc. CONSTITUTION:(A) A DNA cut from a chromosome DNA of Bacillus stearothermophilus MK-232 strain (FERM P-9645) and (B) a competent cell originated from Bacillus subtilis MT-2 strain are subjected to mixed culture to effect halo-formation and obtain (C) a transformant Bacillus subtilis MT-2/ pTZ232. A plasmid DNA pTZ232 (D) is produced by culturing the strain C. The promoter sequence, transcription initiation point, etc., of the component D are searched to separate (E) an SD sequence of formula. The component E is integrated into a manifestation vector such as pUB110 and an E.coli transformed with the resultant plasmid is cultured in a medium to effect the extracellular secretion and production of the desired protein. |
