NEW MATERIAL:A protein, prepared by fusing a protein with a specific binding ability to a lipase and having lipase activity.
USE: A diagnostic agent and biosensor.
PREPARATION: For example, an oligonucleotide complementary to a lipase gene is used to introduce an NcoI site just before a translation starting point and digested with restriction enzymes NcoI and ScaI to separate a DNA fragment containing a lipase gene. Both terminal are then subjected to smoothing treatment and subsequently ligated to a fragment of a plasmid pUC8 treated with the SmaI and then a BAP. The resultant conjugant is further digested with restriction enzymes EcoRI and BamHI to separate a fragment containing the lipase gene. A protein A-fusing expression vector pRIT5 is digested with the NcoI and the terminals are then subjected to smoothing treatment and treated with the EcoRI and BamHI. The aforementioned fragment containing the lipase genge is then ligated to the resultant fragment, further digested with the NcoI and subjected to terminal smoothing treatment. The terminals are then linked to prepare a protein A fusing lipase expression plasmid pAFL-2, which is subsequently inserted into a host. The resultant transformed host is then cultured to afford a protein having lipase activity.
ERIZUMI AKINARI
YOSHINO SHUHEI
INOUE SATOSHI
YOSHIDA NAOYUKI
