PROBLEM TO BE SOLVED: To provide the subject novel shuttle vector that is made impossible of expression of the gene in Escherichia coil by physically separate the control sequence from the structure gene and can restore the gene expression by digestion with the restriction enzyme and following re-cyclization in the Bacillus subtilis. SOLUTION: This novel Bacillus subtilis-Escherichia coil shuttle vector is prepared by physically separating the control sequence from the structure gene and the expression of the gene that is cloned in Escherichia coil is made impossible, while the lost gene expression in the Bacillus substilis is restored by digestion with a kind of restriction enzyme and the following re-cyclization. This is a shuttle vector for mutation and expression and is useful for decomposition of starch, fiber desizing and the production of mutant [alpha]-amylase derives from a bacterium that has excellent merits in other industrial use. This shuttle vector is prepared by treating a wild type [alpha]-amylase gene of Bacillus licheniformis with a chemical to induce a random mutation and linking the mutant body to the vector that can express this mutant.
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|Published - 18-Jul-2000