THEROMOSTABL MUTANT OF SUBSTILISIN J AND ITS PREPARATION METHOD

摘要

A subtilisin J gene of Bacillus stearothermophilus is treated with restriction enzyme Hind II to give DNA segment, introduced to pUC19 and deformed by the mutation-inducing oligonucleotide to obtain the reverse-polymerization chain starting material, followed by polymerization chain reaction(PCR) to give 982bp DNA band which is the first PCR product. The second PCR is performed by adding 17bp forward primer to the 982 bp DNA to obtain precipitate, which is dissolved and treated with restriction enzyme Hind II to give aspartic acid-substituted subtilIsin J.