Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Enzymatic production of a protein or polypeptide
Patent
1987-06-02
1989-01-31
Wiseman, Thomas G.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Enzymatic production of a protein or polypeptide
4351723, 435320, 935 9, 935 12, 935 29, 935 48, C12P 2100, C12N 1500
Patent
active
048015369
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 any polypeptide fragment of flagellin or other proteinaceous material.
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LaVallie Edward R.
Stahl Mark L.
Berstein David L.
Eisen Bruce M.
Genetics Institute Inc.
Mays Thomas D.
Wiseman Thomas G.
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