Cloning, sequencing and expresssion of a gene encoding an eukaryotic amino acid racemase, and diagnostic, therapeutic, and vaccination applications of parasite and viral mitogens
AssigneeInstitut Pasteur
InventorsPaola Minoprio, Mario Arala-Chaves, Antonio Coutinho, Bernardo San Martin, Catherine Rougeot, Wim Degrave, Alain Cosson, Rui Araia Chavas
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Abstract
A method of preventing or inhibiting infection by a parasite or virus in vivo comprises administering to a human in need thereof a parasite or virus mitogen in a sub-mitogenic amount sufficient to induce a protective immune response against the parasite or virus in the human.
Claims32
1-78. (canceled)
A purified nucleic acid molecule
- (a) selected from SEQ ID NOS:
- 8, 9, 10, and 11;
- (b) that encodes a peptide selected from SEQ ID NOS:
- 1, 2, 3, and 4;
- (c) that hybridizes to either strand of a denatured, double-stranded DNA comprising the nucleic acid molecule of (a) or (b) under conditions of moderate stringency;
- (d) derived by in vitro mutagenesis from SEQ ID NOS:
- 8, 9, 10, and 11;
- (e) degenerate from SEQ ID NOS:
- 8, 9, 10, and 11 as a result of the genetic code;
- (f) that encodes Tc45 polypeptide, an allelic variant of Tc45 polypeptide, or a homolog of Tc45 polypeptide;
- (g) that encodes an eukaryotic protein with an amino acid racemase activity;
- (h) that encodes an eukaryotic protein with a proline racemase activity;
- (i) that encodes an eukaryotic protein, which is recognized by antibodies raised against an eukaryotic protein having proline racemase activity;
- (j) that has at least 80% of identity with the sequence of an eukaryotic gene encoding a protein with a racemase activity;
- and/or
- (k) that is a fragment of a polynucleotide containing at least 50 nucleotides of the sequence of the proline racemase gene of T. cruzi or hybridizing under stringent conditions with a polynucleotide according to any one of (g), (h), (i) or (j).
A process for screening a molecule capable of inhibiting the amino acid racemase activity of a eukaryotic protein comprising:
- (a) contacting the purified eukaryotic racemase protein with standard doses of a molecule to be tested;
- (b) measuring inhibition of racemase activity;
- and
- (c) selecting the molecule.
A method for the production of D-amino acid using a purified eukaryotic amino acid racemase comprising:
- (a) incubating L-amino acid with the recombinant eukaryotic amino acid racemase;
- (b) separating the D-amino acid produced in (a);
- and
- (c) purifying the D-amino acid.
A method of preventing or inhibiting infection by a virus or a protozoan parasite in vivo, wherein the method comprises administering to a subject in need thereof a virus or a protozoan parasite mitogen in a sub-mitogenic amount sufficient to induce a protective immune response against the virus or the protozoan parasite in the subject, wherein the virus or protozoan parasite mitogen is optionally administered to the subject in admixture with a pharmaceutically acceptable carrier.
Dependencies are inferred from the claim text.