FOREIGN GENE EXPRESSION AND SECRETION SYSTEM IN THE GENUS BIFIDOBACTERIUM TO PRODUCE FUNCTIONAL LACTIC ACID BACTERIA PRODUCT PRODUCING USEFUL PROTEIN

摘要

PURPOSE: A foreign gene expression and secretion system in the genus Bifidobacterium is provided, thereby producing functional lactic acid bacteria product producing useful protein, and applying the expression cassette system containing alpha-amylase signal sequence from Bifidobacterium into industrial system. CONSTITUTION: A alpha-amylase gene from Bifidobacterium having the nucleotide sequence set forth in SEQ ID NO:1 is provided. A signal sequence of the alpha-amylase gene from Bifidobacterium having the nucleotide sequence set forth in SEQ ID NO:2 is provided. A cassette system for secretion of foreign gene contains the signal sequence of the alpha-amylase gene from Bifidobacterium. A promoter gene of alpha-amylase gene from Bifidobacterium has the nucleotide sequence of SEQ ID NO:3. An expression vector of pBESAF1 or pBESAF2(KCCM-10452) comprises the promoter gene of alpha-amylase gene from Bifidobacterium having the nucleotide sequence of SEQ ID NO:3, ribosome binding site and the signal sequence of the alpha-amylase gene from Bifidobacterium having the nucleotide sequence of SEQ ID NO:2. A phytase expression vector of pBESAF1-PHY or pBESAF2-appA comprises phytase gene from Bacillus and Escherichia coli. A E. coli or Bifidobacterium transformant is produced by transforming E. coli or Bifidobacterium with the expression vector of pBESAF1 or pBESAF2(KCCM-10452), or the phytase expression vector of pBESAF1-PHY or pBESAF2-appA.