Gene expression technique

摘要

The present invention provides a method for producing a desired protein (such as a desired heterologous protein) comprising: (a) providing a host cell comprising a first recombinant gene encoding a protein comprising the sequence of a first chaperone protein, a second recombinant gene encoding a protein comprising the sequence of a second chaperone protein and a third gene, such as a third recombinant gene, encoding a desired protein (such as a desired heterologous protein), wherein the first and second chaperones are different; and (b) culturing the host cell in a culture medium to obtain expression of the first, second and third genes.

主权项

1. A plasmid comprising a gene encoding a heterologous protein, and as the sole yeast selectable marker, one or more genes encoding an essential chaperone that is essential to the viability of a yeast host cell when the one or more genes encoding the essential chaperone are deleted or inactivated in a yeast host cell and the host cell is inviable in culture and the deficiency cannot be complemented by additions or modifications to the culture medium, wherein the natural location of the one or more genes encoding the essential chaperone are modified to disrupt its function under normal growth conditions wherein the essential chaperone is a yeast chaperone comprising the sequence of a protein encoded by a gene selected from the group consisting of Protein Disulfide Isomerase (PDI) and Protein Disulfide Isomerase 1(PDI1).