HETEROLOGOUS PROTEIN EXPRESSION IN NEUROSPORA

THE PROPOSED RESEARCH WILL INVESTIGATE THE ABILITY OF NEUROSPORA CRASSA TO EXPRESS AND SECRETE BIOLOGICALLY-ACTIVEMAMMALIAN PROTEINS. THE ABILITY TO GENETICALLY ENGINEER BACTERIA AND YEASTS FOR THESE TASKS IS WELL DOCUMENTED. HOWEVER, RECOMBINANT PRODUCTS FROM THESE ORGANISMS OFTEN REQUIRE COSTLY DOWNSTREAM PURIFICATION AND PROCESSING. THE PHYSIOLOGY OF THE FILAMENTOUS FUNGI INDICATE THAT THEY ARE EXCELLENT MICROBES TO EFFICIENTLY EXPRESS, MODIFY, AND SECRETE HETEROLOGOUS PROTEINS. SOME INDUSTRIAL FUNGAL STRAINS SECRETE ENDOGENOUS ENZYMES IN MILLIGRAM PER LITER QUANTITIES. THUS, THE INEXPENSIVELY CULTURED FILAMENTOUS FUNGI PROVIDE THE OPPORTUNITY TO SIGNIFICANTLY REDUCE THE COST OF PRODUCING PHARMACOLOGICALLY ACTIVE PROTEINS BY MICROORGANISMS. THE PROPOSED RESEARCH WILL PLACE CDNAS ENCODING RAT RELAXIN AND BOVINE CHYMOSIN UNDER THE REGULATION OF THE N.CRASSA BETA-TUBULIN GENE PROMOTER AND TERMINATOR, AND TEST THE ABILITY OF NEUROSPORA TO EXPRESS THE RECOMBINANT GENES AND TO SECRETE THE HETEROLOGOUS PROTEINS USING THE HETEROLOGOUS PROTEINS' OWN SECRETION SIGNAL PEPTIDES.