Leland Stanford Junior University

Robert T Schimke

Stanley N Cohen

Jack H Nunberg

Annie C Chang

Susan M Thorpe

John M Abrams

Methods and compositions are provided for producing heterologous proteins in a micrcorganism host, particularly bacterium, without fusion to an endogenous protein. Particularly, the heterologous gene may be inserted into a vector comprising an endogenous gene, where the heterologous gene is preceded by a ribosomal binding site. A heterologous enzyme functional in a bacterial host is demonstrated.

Protein production at synthetic start site

DNA constructs are provided employing intronically positioned expression, systems, where one of the genes is a dominant gene, usually amplifiable, and the other gene encodes a sequence of interest. Higher levels of co-expression are achieved than when the genese are ligated in tandem. Specifically, the gene of interest is inserted into the intron of a DHFR minigene, the construct transformed into a mammalian cell and the resulting transformants stressed with progressively increasing levels of methotrexate. Substantially increasing levels of co-expression are achieved with increasing levels of methotrexate.

Growing community of inventors

Palo Alto, CA, United States of America

Robert T Schimke