CLONING, SEQUENCING, AND EXPRESSION OF HUMAN BSA LIPASE CDNA

BILE SALT-ACTIVATED LIPASE (BSAL) IS FOUND IN THE MILK OF A LIMITED NUMBER OF MAMMALS: HUMANS, GORILLAS, DOGS, AND CATS THE ENZYME PLAYS AN IMPORTANT ROLE IN THE INFANT DIET IN THEDIGESTION OF LIPIDS, CAUSING A STRIKING QUANTITATIVE DIF- FERENCE BETWEEN BREAST-FED AND FORMULA-FED INFANTS IN WEIGHTGAIN. IN THIS REGARD, IT IS IMPORTANT TO NOTE THAT BSAL IS ABSENT IN BOVINE MILK. IN PHASE I, ZYMOGENETICS, INC, PLANS TO CLONE, SEQUENCE, ANDEXPRESS THE CDNA FOR HUMAN BSAL. THIS WILL PROVIDE A FRAME WORK FOR PHASE II RESEARCH, WHEREIN RECOMBINANT HUMAN BSAL WILL BE PRODUCED IN LARGE QUANTITIES FOR SUPPLEMENTATION IN INFANT FORMULAS. EXPRESSION OF THE HUMAN BSAL CDNA WILL BE ACHIEVED BY CLONING INTO AN EXPRESSION VECTOR SUITABLE FOR PRODUCTION AT HIGH LEVELS IN SACCHAROMYCES CEREVISIAE. LEVELS OF EXPRESSION WILL BE ASCERTAINED BY ELISA AND DIRECTMEASUREMENTS OF ENZYMATIC ACTIVITY. THE RECOMBINANT PROTEINWILL BE PURIFIED AND CHARACTERIZED FOR ITS PHYSICAL AND CHEMICAL SIMILARITY TO THE PURIFIED HUMAN MILK ENZYME