Method for producing heterologous proteins

US4801536A · kind A · utility

Assignee

• Genetics Institute, LLC · US

Inventors

• Mark Stahl · Lexington, US
• Edward R. LaVallie · Harvard, US

Key dates

• Filing date: Jun 2, 1987
• Grant date: Jan 31, 1989
• Expiry date: Jun 2, 2007

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C07K2319/75
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

This invention concerns a method for producing a heterologous protein in a bacterial host cell such that the protein is exported from the host cell into the culture medium. The method involves culturing in a bacterial culture medium a genetically engineered bacterial strain containing a fusion DNA sequence comprising a first nucleotide sequence encoding at least an N-terminal portion of a flagellin protein and a second nucleotide sequence encoding the heterologous protein. The first nucleotide sequence is linked via its 3' terminus to the 5' terminus of the second nucleotide sequence, and the fusion DNA sequence is itself linked to an expression control sequence. In certain embodiments the first and second nucleotide sequences are linked by means of a linking nucleotide sequence encoding a selectively cleavable polypeptide, In those embodiments the resulting exported fusion protein will contain a selectively cleavable site at which the fusion protein may be selectively cleaved by chemical or enzymatic methods to produce the heterologous protein encoded for by the second nucleotide sequence of the fusion DNA sequence. The heterologous protein may then be separately recovered from an…