METHOD FOR SECRETING AND PRODUCING PROTEIN WITH ESCHERICHIA COLI

摘要

PURPOSE:To obtain a recombinant expression secretory plasmid, comprising a base sequence containing a promoter derived from a specific microorganism, a ribosome binding site and a region capable of coding a secretion signal, a desired proteinic structural gene and a replication origin and capable of highly secreting a useful protein. CONSTITUTION:A base sequence containing a promoter derived from Bacillus.amyloliquefaciens obtained from a plasmid pNPA225, a ribosome binding site and a region capable of coding a secretion signal is linked to a base sequence excised from a plasmid pHGH10 containing a structural gene capable of coding a desired protein (e.g. human growth hormone) to synthesize a plasmid pNPG5, which is then linked to a plasmid pUC19 derived from Escherichia coli as a replication origin for replication in Escherichia coli to synthesize a recombinant plasmid pHGE195. The resultant plasmid is subsequently transduced into the Escherichia coli to carry out transformation. The obtained transformant is then cultured to efficiently afford an extraneous useful protein (e.g. the human growth hormone).