WO2004005489A2 - Acides nucleiques codant l'antigene sarcosystis neurona et leurs utilisations - Google Patents

Acides nucleiques codant l'antigene sarcosystis neurona et leurs utilisations Download PDF

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Publication number
WO2004005489A2
WO2004005489A2 PCT/US2003/004856 US0304856W WO2004005489A2 WO 2004005489 A2 WO2004005489 A2 WO 2004005489A2 US 0304856 W US0304856 W US 0304856W WO 2004005489 A2 WO2004005489 A2 WO 2004005489A2
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Prior art keywords
seq
neurona
nucleic acid
set forth
antigen
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PCT/US2003/004856
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English (en)
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WO2004005489A3 (fr
Inventor
Daniel K. Howe
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University Of Kentucky Research Foundation
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Priority to AU2003217568A priority Critical patent/AU2003217568A1/en
Publication of WO2004005489A2 publication Critical patent/WO2004005489A2/fr
Publication of WO2004005489A3 publication Critical patent/WO2004005489A3/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/44—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from protozoa
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the present Invention relates to nucleic acids of Sarcocystis
  • the present invention relates to nucleic acids of
  • the present invention relates to novel
  • primers including primers, probes, antigen/antibody diagnostic kits, vectors for
  • Sarcocystis neurona is an apicomplexan parasite that is the
  • EPM equine protozoal myeloencephalitis
  • EPM is a common and debilitating infectious disease that affects
  • S.. neurona (Dubey et al., 1991). S. neurona is related to the human and
  • Neospora spp. pathogen Neospora spp. These species are phylogenetically classified
  • Virginians serving as a definitive host (Fenger et al, 1995).
  • intermediate host(s) include skunks (Cheadle et al, 2001b), raccoons
  • felids may be only an experimental
  • S. neurona detection have been developed based on the S. neurona
  • PCR-based assays detect the presence of S. neurona DNA
  • present invention hereby allow for the production of recombinant
  • antibodies provided by the invention serve as valuable reagents for
  • a recombinant S. neurona antigen furnishes the key component for a
  • parasite lysate provides a second-generation assay that significantly
  • Thl two arms of the immune system are characterized by Thl or Th2
  • lymphocytes that differ in their profile of secreted cytokines, and these
  • Immunoglobulin isotype switching is an important
  • Thl profile is skewed towards a Thl profile will be characterized by IgG2a and
  • IgG3 (Finkelman et al, 1990; Snapper et al, 1997). It is generally
  • Thl cell-mediated response is necessary for control of
  • neurona infection is unclear but may be secondary or unimportant.
  • antigen for development of a diagnostic test can be somewhat subjective since any particular pathogen is composed of numerous
  • antigens of the Coccidia are exceedingly immunogenic.
  • antigens of the Coccidia are exceedingly immunogenic.
  • Neospora caninum Remington, 1980; Sharma et al, 1983
  • Neospora caninum Howe
  • SAGs SAG-related sequences
  • SRSs SAG-related sequences
  • antigens are involved in modulation of the host immune response
  • gondii has been shown to protect mice against acute toxoplasmosis
  • S. neurona i.e., SnSAG2, SnSAG3,
  • the present invention utilizes recombinant S. neurona
  • Toxoplasma gondii SAGl induces protective cells into both NALT and
  • gondii consisting of 824-nucleotides encoding the 275 amino acid
  • mice immunized with a plasmid encoding the SAGl gene mice immunized with a plasmid encoding the SAGl gene. Infection and Immunity, 1999, 67: 6358-6363.
  • Peterson et al. discloses the use of an E. coli produced vaccine
  • E. coli produced recombinant T gondii SAGl with alum as adjuvant
  • mice protect mice against lethal infection with Toxoplasma gondii. Vaccine.
  • Bonieri et al. discloses intranasal immunity with SAGl and
  • Haumont et al. discloses that a recombinant form of Toxoplasma
  • Angus et al. discloses that immunization with a DNA plasmid
  • mice Sera of immunized mice showed recognition of T. gondii tachyzoites by immunofluorescence and
  • nucleic acid vaccination can provide protection against T. gondii
  • mice infection in mice. See, Angus CW, Klivington-Evans D, Dubey JP,
  • neurona merozoite culture that is chemically inactivated
  • neurona In particular, it is an object of the present invention to
  • nucleic acids capable of selectively hybridizing with the nucleic acid
  • Sarcocystis neurona including, but not limited to, primers and
  • Another object of the invention is to provide a vector comprising
  • One important object of the invention is to provide a purified
  • a particular object of the present invention is to
  • Sarcocystis neurona and a method of detection of Sarcocystis neurona utilizing the antibodies of the present invention.
  • the present invention satisfies the need in the art by providing a
  • Sarcocystis neurona or a unique fragment thereof.
  • Sarcocystis neurona or a unique fragment thereof.
  • the invention provides novel isolated nucleic acids encoding membrane-
  • the present invention also provides purified antigenic
  • the invention provides purified antigenic proteins or purified antigenic
  • the present invention provides a purified antigenic
  • the present invention also provides isolated nucleic acids
  • Sarcocystis neurona including, but not limited to, primers and probes
  • PCR polymerase chain reaction
  • the present invention provides vectors comprising the
  • the present invention also provides a purified polyclonal
  • Figure 1 is a sequence comparison of SnSAGl, SnSAG3, and
  • SnSAG4 with TgSAG2E.
  • the S. neurona surface antigens SnSAGl, SnSAG3 and SnSAG4 are most similar to the TgSAG2 family of T.
  • SnSAGs contain 10/12 conserved cysteine residues that have been
  • Figure 2 is a sequence comparison of SnSAG2 with TgSAG 1
  • the S. neurona surface antigen SnSAG2 is most similar
  • SnSAG2 shares modest sequence identity to its TgSAG
  • SnSAG2 will also align with the carboxyl-terminal domain of the
  • Figure 3 shows a Western blot analysis of the Sn SAGs in S.
  • the SnSAG genes were expressed in E. coli, and
  • SnSAG2 corresponded to an immunodominant band at approximately
  • FIG. 4 shows the SnSAGs are membrane-associated in
  • FIG. 5 shows that the four SnSAGs are displayed on the
  • the present invention satisfies the long felt need in the art by
  • nucleic acid means a chain of at least two or more nucleotides such as DNA
  • RNA ribonucleic acid
  • purified nucleic acid is one that is substantially separated from other
  • isolated nucleic acid is meant
  • nucleic acids of the present invention are separated from at least some of other nucleic acids found in the naturally-occurring organism.
  • RNA can include positive and negative strand RNA as well as DNA.
  • RNA and RNA are meant to include genomic and subgenomic nucleic acids
  • nucleic acids contemplated by the present invention include a
  • nucleic acid having sequences from which a Sarcocystis neurona cDNA
  • unique fragment is meant a fragment of the nucleic acids set forth in
  • the invention are also contemplated as long as the essential structure and function of the polypeptide encoded by the nucleic acids is
  • fragments used as primers or probes can have
  • nucleic acids of the invention are the nucleic acids of the invention.
  • homologs or naturally are naturally occurring amino acids of the invention.
  • invention can range from about 50% to about 99% sequence identity to
  • one embodiment of the present invention provides
  • polypeptide is meant a polypeptide that has been substantially separated
  • Sarcocystis neurona is an apicomplexan parasite that can cause a
  • EPM myeloencephalitis
  • S. neurona is an obligate intracellular pathogen that
  • Parasite surface molecules are virulence factors that are typically novel and undoubtedly important
  • TgSAG2 TgSAG2.
  • the present invention provides identity and
  • the present invention provides four isolated nucleic acids of
  • sequence database has revealed a family of at least four S. neurona
  • T gondii the novel & neurona surface antigens in T gondii. Based on their homology to the T. gondii SAGs, the novel & neurona surface antigens have been designated SnSAGl,
  • SnSAGl proteins have been designated SnSAGl, SnSAG2, SnSAG3, and
  • one embodiment of the present invention comprises
  • the nucleic acid identified in SEQ ID NO: 21 comprises an
  • neurona which encodes a 276 amino acid polypeptide set forth in the Sequence Listing as SEQ ID NO: 22.
  • the polypeptide encoded by SEQ ID NO: 22 The polypeptide encoded by SEQ ID NO: 22.
  • ID NO: 22 has a predicted amino-terminal signal peptide (indicating
  • CSF cerebrospinal fluid
  • the nucleic acid identified in SEQ ID NO: 23 comprises an 975
  • neurona which encodes a 168 amino acid polypeptide set forth in the Sequence Listing as SEQ ID NO: 24.
  • the present invention also provides an isolated nucleic acid as
  • SEQ ID NO: 25 comprises an 1585 nucleotide open
  • nucleic acid Also provided by the present invention is an isolated nucleic acid
  • SEQ ID NO: 27 comprises an 1111 nucleotide open
  • the recombinant proteins provided by the invention can be used as reagents
  • sequence tags generated from the cSn.l cDNA library has
  • SnGFl S. neurona Gene
  • SnGFl encodes a set of similar proteins (at least eight)
  • SnGFl a-h SnGFl a-h. These genes are predicted to encode proteins of,
  • These proteins have a predicted N-terminal signal peptide and a
  • one embodiment of the present invention provides
  • SnGFl a an isolated nucleic acid designated SnGFl a which comprises the
  • Another embodiment of the invention comprises the polypeptide sequence encoded by SnGFl a set forth in the Sequence
  • SnGFlb isolated nucleic acid designated SnGFlb which comprises the nucleic acid
  • SnGFl c isolated nucleic acid designated SnGFl c which comprises the nucleic
  • SnGFld isolated nucleic acid designated SnGFld which comprises the nucleic acid
  • Another embodiment of the invention comprises the polypeptide
  • the present invention also provides an isolated nucleic acid
  • SnGFl e which comprises the nucleic acid set forth in SEQ
  • SnGFlf isolated nucleic acid designated SnGFlf which comprises the nucleic acid
  • SnGFl g isolated nucleic acid designated SnGFl g which comprises the nucleic
  • Another embodiment of the invention comprises the polypeptide
  • SnGFlh isolated nucleic acid designated SnGFlh which comprises the nucleic acid
  • the present invention provides isolated nucleic acids as set forth
  • Purified polypeptides encoded by the nucleic acids are also provided.
  • polypeptides can be utilized in methods of diagnosis or as
  • Vectors are also provided.
  • nucleic acids of the present invention are provided which comprise the nucleic acids of the present invention.
  • vectors can be utilized in host expression systems to produce antigenic
  • the present invention also provides purified antibodies selectively reactive
  • the invention provides purified antigenic
  • the invention also provides these antigenic polypeptides in a
  • polypeptides can be deduced from the nucleotide sequences set forth in
  • purified means the antigen is at least sufficiently free of contaminants
  • the present invention are also referred to herein as "the antigen" or "the
  • antigenic fragments can be any antigenic fragments. It is contemplated that the antigenic fragments can be any antigenic fragments.
  • one example provides an
  • An antigenic fragment of the antigen can be isolated from the
  • the antigen can also be synthesized directly.
  • An immunoreactive substance an immunoreactive substance
  • fragment is generally an amino acid sequence of at least about five amino acids
  • polypeptide fragments of the present invention can also be any polypeptide fragments of the present invention.
  • polypeptide or fragments thereof Once the amino acid sequence of the antigen is provided, it is provided.
  • amino acid sequences of the present polypeptides can be any amino acid sequences of the present polypeptides.
  • amino acid sequences of an S. neurona antigen can be any amino acid sequences of an S. neurona antigen.
  • the peptide should be any case, the peptide should
  • bioactive property such as immunoreactivity, immunogenicity,
  • antigen administered depend on the subject, e.g. a horse or a guinea pig,
  • animal so inoculated with the antigen can be exposed to the parasite to
  • a vector comprising the nucleic acids of the present invention is
  • the vectors of the invention can be in a host capable of
  • microbial hosts suitable for use include bacilli, such as Bacillus subtilis, and other enterobacteriaceae, such as Salmonella,
  • promoters such as the lactose promoter
  • Trp tryptophan
  • ribosome binding site sequences for example, for initiating and
  • amino terminal methionine can be provided by insertion of a Met codon
  • yeast expression can be used.
  • yeast secretory systems In one example, the Saccharomyces
  • cerevisiae pre-pro-alpha-factor leader region encoded by the
  • MF.alpha.-l gene is routinely used to direct protein secretion from
  • this enzyme cleaves the precursor protein on the carboxyl side of a
  • sequence can be fused in-frame to the pre-pro-alpha-factor leader
  • This construct is then put under the control of a strong
  • transcription promoter such as the alcohol dehydrogenase I promoter or
  • the antigen coding sequence is followed by a
  • antigen coding sequences can be any antigen coding sequence.
  • antigen coding sequences can be any antigen coding sequence.
  • .beta.-galactosidase used to facilitate purification of the fusion protein
  • Mammalian cells permit the expression of proteins in an
  • the vectors can contain genes conferring either
  • gentamicin or methotrexate resistance for use as selectable markers.
  • the antigen and immunoreactive fragment coding sequence can be any suitable antigen and immunoreactive fragment coding sequence.
  • CHO cell lines include the CHO cell lines, HeLa cells, myeloma cell lines,
  • Expression vectors for these cells can include
  • expression control sequences such as an origin of replication, a
  • RNA splice sites as ribosome binding sites, RNA splice sites, polyadenylation sites, and transcriptional terminator sequences.
  • sequences are promoters derived from immunoglobulin genes, SV40,
  • nucleic acid segments of interest can be transferred into the host cell by
  • prokaryotic cells prokaryotic cells, whereas calcium phosphate treatment or
  • electroporation may be used for other celluar hosts.
  • tissue plasminogen activator gammainterferon, tissue plasminogen activator, clotting Factor VIII,
  • the vector can include CMV
  • the nucleic acid sequences can be expressed in hosts after the
  • sequences have been operably linked to, i.e., positioned to ensure the
  • expression vectors can contain selection markers, e.g., tetracycline
  • Polynucleotides encoding a variant polypeptide may include
  • polynucleotides can include a
  • eukaryotic expression hosts eukaryotic expression hosts
  • a ribosome binding site eukaryotic expression hosts
  • a ribosome binding site eukaryotic expression hosts
  • an enhancer for use in eukaryotic expression hosts and, optionally,
  • peptides of the invention comprises the use of Alphavirus vector
  • the antibodies can be specifically reactive
  • reactive means capable of binding or
  • Antibodies can be made as described
  • Antibodies can either be purified directly, or spleen cells can be
  • the antibodies can be any antibodies that can be used for antibody secretion.
  • the antibodies can be any antibodies that can be used for antibody secretion.
  • detectable moiety or both bound and labeled.
  • contemplated by the present invention include, but are not limited to
  • purified ligand specifically reactive with the antigen can be an antibody.
  • the antibody can be a monoclonal antibody obtained by standard
  • the monoclonal antibody can be any monoclonal antibody.
  • the polyclonal antibody can also be obtained by the standard
  • the present invention provides a method of detecting the
  • One example of the method of detecting S. neurona is
  • the antigen will be on intact cells
  • the antibody includes any ligand which binds the
  • antigen for example, an intact antibody, a fragment of an antibody or
  • this method can comprise any body fluid which would contain the
  • antigen or a cell containing the antigen such as blood, plasma, serum, cerebrospinal fluid, saliva, feces and urine.
  • blood, plasma, serum, cerebrospinal fluid, saliva, feces and urine Other possible examples of
  • body fluids include sputum, mucus, gastric juice and the like.
  • Enzyme immunoassays such as immunofluorescence assays
  • IF A enzyme linked immunosorbent assays (ELISA) and
  • immunoblotting can be readily adapted to accomplish the detection of
  • antigen can, for example, be as follows: (1) bind the antibody to a
  • detectable moiety e.g., horseradish peroxidase enzyme or
  • MAbs monoclonal antibodies
  • binding is measured relative to a control (no patient semm antibody).
  • the degree of monoclonal antibody inhibition is a very specific test for
  • MAbs can also be used for detection directly in cells
  • a micro-agglutination test can also be used to detect the
  • latex beads or red blood
  • the agglutinated antigen-antibody complexes form a
  • the antibody can be any substance that can be used as in a typical sandwich assay.
  • the antibody can be any substance that can be used as in a typical sandwich assay.
  • invention provides S. neurona antigen for the detection of infectious, S.
  • the antigen can be any substance that can be used in the diagnostic methods taught herein.
  • the antigen can be any substance that can be used in the diagnostic methods taught herein.
  • a substrate bound to a substrate and contacted by a fluid sample such as serum
  • antibody bound to, or labeled with, a detectable moiety can be added to
  • the secondary antibody or other ligand which is reactive either specifically
  • ligand or reacted antibody will be selected for its ability to react with
  • the detectable moiety will allow visual detection of a precipitate
  • detectable moieties include fluorescein and rhodamine (for
  • biotin-streptavidin for example
  • alkaline phosphatase for biochemical
  • the antigen e.g., a purified antigenic polypeptide fragment
  • the vaccine can be any suitable pharmaceutically acceptable carrier.
  • the vaccine can be any suitable pharmaceutically acceptable carrier.
  • the vaccine can be any suitable pharmaceutically acceptable carrier.
  • the vaccine can be any suitable vaccine, or other strain, or an epitope specific to the antigen.
  • the vaccine can be any suitable vaccine, or other strain, or an epitope specific to the antigen.
  • Immunogenic amounts of the antigen can be determined using
  • the pharmaceutically acceptable carrier can comprise saline or
  • An adjuvant can also be a
  • the mode of administration is part of the carrier of the vaccine, in which case it can be selected by standard criteria based on the antigen used, the mode of administration
  • the invention as a prophylactic or a therapeutic modality.
  • the antigenic agent for use in the vaccines of the invention is selected from the group consisting of the amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids, amino acids
  • inventions can be any nucleic acid, e.g., as set forth in the Sequence
  • acids include those that encode the native proteins of S. neurona, e.g.,
  • the nucleic acid used as a vaccine can be e.g. , a naked DNA, or the nucleic acid can be incorporated in an expression vector as set forth
  • the presence of S. neurona can also be determined by detecting
  • invention provides a method of detecting the presence of S. neurona in
  • a subject comprising detecting the presence of the nucleic acid
  • the nucleic acid specific for S. neurona can be detected utilizing
  • nucleic acid amplification technique such as polymerase chain
  • nucleic acid is detected utilizing direct hybridization or by utilizing a restriction
  • the presence of amplification indicates the presence of S.
  • nucleic acid sample can be sequenced directly using, techniques known
  • invention provides a method of detecting the presence of S. neurona by
  • embodiment S. neurona can be detected by directly hybridizing the
  • nucleotide sequence could be amplified prior to
  • LCR LCR
  • mismatch probes i.e., probes which are fully complementary with the target except at the point of the mutation.
  • target sequence is then allowed to hybridize both with oligonucleotides
  • PCR polymerase chain reaction
  • reverse transcriptase reverse transcriptase
  • PCR are techniques that amplify specific nucleic acid sequences with
  • polymerase e.g., a heat stable
  • oligonucleotides can be prepared which are complementary to
  • oligonucleotide is complementary to one of the two strands.
  • oligonucleotides oriented with their 3' ends pointing towards each
  • the end product is
  • PCR may be followed by restriction
  • endonuclease recognition site facilitates the detection of the organism
  • RFLP restriction fragment length polymorphism
  • nucleic acid is obtained, for example from
  • S. neurona nucleic acid is detected and their mobility on the gel by
  • SSCA sequence analysis
  • primers for PCR and LCR are usually about 20 bp in
  • Denaturation of strands usually takes place at about 94.degree. C. and extension from the primers is usually at
  • the annealing temperature varies according to the
  • reaction times 20 mins
  • PASA specific alleles
  • allele specific amplification involves amplification with two
  • oligonucleotide primers such that one is allele-specific.
  • LCR ligase chain reaction
  • LCR probes may be combined or multiplexed
  • LCR can be particularly useful where, as here, multiple mutations are
  • the present invention is more particularly described in the
  • SnSAGl surface antigen gene
  • SnSAGl recognized a 25-kDa antigen in western blots of non-reduced
  • S. neurona indicated that the protein is expressed throughout

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Abstract

L'invention concerne de nouveaux acides nucléiques isolés codant des protéines antigènes dérivées de Sarcosystis neurona, ou des fragments uniques de celles-ci. En particulier, l'invention concerne de nouveaux acides nucléiques isolés codant des polypeptides associés aux membranes SnSAG2, SnSAG3, et SnSAG 4. L'invention concerne également des fragments polypeptidiques antigènes purifiés codés par ces nouvelles séquences d'acides nucléiques qui codent Sarcosystis neurona. En particulier, l'invention concerne des fragments polypeptidiques antigènes purifiés codés par ces nouvelles séquences d'acides nucléiques qui codent pour SnSAG2, SnSAG3, et SnSAG 4. En outre, l'invention concerne un fragment polypeptidique antigène purifié codé par les séquences d'acides nucléiques ou une partie choisie de celles-ci, dans un porteur pharmaceutiquement acceptable. Elle concerne également des acide nucléiques isolés capables de s'hybrider de manière sélective avec l'acide nucléique provenant de Sarcosystis neurona. Elle concerne en outre des vecteurs comprenant lesdits acides nucléiques codant Sarcosystis neurona ou un fragment de celui-ci, ainsi que le vecteur d'un hôte capable d'exprimer le polypeptide codé par l'acide nucléique en question. Enfin, l'invention concerne un anticorps polyclonal et/ou monclonal purifié qui réagit spécifiquement avec Sarcosystis neurona, ainsi qu'un procédé permettant de détecter Sarcosystis neurona utilisant ces anticorps.
PCT/US2003/004856 2002-02-15 2003-02-19 Acides nucleiques codant l'antigene sarcosystis neurona et leurs utilisations WO2004005489A2 (fr)

Priority Applications (1)

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AU2003217568A AU2003217568A1 (en) 2002-02-15 2003-02-19 Nucleic acids encoding sarcocystic neurona antigen and uses thereof

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US35747902P 2002-02-15 2002-02-15
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Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
DATABASE GENBANK [Online] 'Sarcocystis neurona EST project', XP002904286 Database accession no. (BM252380) *

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