Methods of using a Mycobacterium tuberculosis coding sequence to facilitate stable and high yield expression of the heterologous proteins
AssigneeCorixa Corporation
InventorsYasir Skeiky, Jeffrey Guderian
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Abstract
The present invention relates generally to nucleic acid and amino acid sequences of a fusion polypeptide comprising a Mycobacterium tuberculosis polypeptide, and a heterologous polypeptide of interest, expression vectors and host cells comprising such nucleic acids, and methods for producing such fusion polypeptides. In particular, the invention relates to materials and methods of using such M. tuberculosis sequence as a fusion partner to facilitate the stable and high yield expression of recombinant heterologous polypeptides of both eukaryotic and prokaryotic origin.
Claims24
A recombinant nucleic acid molecule that encodes a fusion polypeptide, the recombinant nucleic acid molecule comprising a Ra12 polynucleotide sequence and a non-Mycobacterium tuberculosis polynucleotide sequence, wherein the Ra12 polynucleotide sequence encodes a Ra12 polypeptide consisting of the sequence set forth in SEQ ID NO:4, SEQ ID NO:17, SEQ ID NO:18, or SEQ ID NO:23.
A recombinant nucleic acid molecule that encodes a fusion polypeptide, the recombinant nucleic acid molecule comprising a Ra12 polynucleotide sequence and a non-Mycobacterium tuberculosis polynucleotide sequence, wherein the Ra12 polynucleotide sequence encodes a Ra12 polypeptide consisting of the sequence set forth in SEQ ID NO:17.
A recombinant nucleic acid molecule that encodes a fusion polypeptide, the recombinant nucleic acid molecule comprising a Ra12 polynucleotide sequence and a non-Mycobacterium tuberculosis polynucleotide sequence, wherein the Ra12 polynucleotide sequence encodes a Ra12 polypeptide consisting of the sequence set forth in SEQ ID NO:18.
A recombinant nucleic acid molecule that encodes a fusion polypeptide, the recombinant nucleic acid molecule comprising a Ra12 polynucleotide sequence and a non-Mycobacterium tuberculosis polynucleotide sequence, wherein the Ra12 polynucleotide sequence encodes a Ra12 polypeptide consisting of the sequence set forth in SEQ ID NO:4.
A method of producing a fusion polypeptide, the method comprising:
- expressing in a host cell a recombinant nucleic acid molecule that encodes a fusion polypeptide, the fusion polypeptide comprising a Ra12 polypeptide and a non-Mycobacterium tuberculosis polypeptide, wherein the Ra12 polypeptide consists of the sequence set forth in SEQ ID NO:4, SEQ ID NO:17, SEQ ID NO:18, or SEQ ID NO:23;
- and
- purifying the fusion polypeptide from the host cell.
A recombinant nucleic acid molecule that encodes a fusion polypeptide, the recombinant nucleic acid molecule comprising a Ra12 polynucleotide sequence and a non-Mycobacterium tuberculosis polynucleotide sequence, wherein the Ra12 polynucleotide sequence encodes a Ra12 polypeptide consisting of the sequence set forth in SEQ ID NO:23.
Dependencies are inferred from the claim text.