Methods and compositions for the detection of bovine pregnancy which utilize members of the 1-8 family of interferon inducible genes
OwnerThomas R. Hansen, Kathleen J. Austin +1inventors
InventorsThomas R. Hansen, Kathleen J. Austin, James K. Pru
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Abstract
Provided herein are two 1-8 family gene interferon tau inducible genes, bovine 1-8U and bovine Leu-13, and methods of detecting the same to determine bovine pregnancy.
Claims35
An isolated double-stranded nucleic acid molecule which upon denaturation, specifically hybridizes with SEQ ID NO 1, said nucleic acid molecule comprising a sequence encoding a bovine 1-8U protein of about 16 Kda, the expression of said protein being induced by interferon and elevated during bovine pregnancy when compared to levels in non-pregnant bovine animals.
An isolated nucleic acid molecule comprising a sequence selected from the group consisting of:
- a) SEQ ID NO 1;
- b) a sequence which specifically hybridizes with SEQ ID NO 1;
- and
- c) a sequence encoding a polypeptide of SEQ ID NO 2.
An oligonucleotide between about 10 and about 200 nucleotides in length, which specifically hybridizes with a protein translation initiation site in a nucleotide sequence encoding amino acids of SEQ ID NO 2.
An isolated bovine 1-8U protein, about 16 kDa encoded by SEQ ID NO:
- 2, expression of said encoded protein being induced by interferon and elevated during bovine pregnancy.
A nucleic acid comprising the 5′ untranslated, promoter region of a bovine 1-8U gene, comprising SEQ ID NO:
- 11.
A method for identifying agents which modulate bovine 1-8U promoter activity comprising:
- a) introducing a nucleic acid construct comprising the bovine 1-8U promoter region operably linked to a reporter gene into a host cell;
- b) contacting said cell with an agent suspected of modulating bovine 1-8U promoter activity;
- and
- c) determining the ability of said agent to modulate said promoter activity as a function of reporter gene expression levels relative to control cells comprising said construct which are not contacted with said agent.
An isolated double-stranded nucleic acid molecule which upon denaturation, specifically hybridizes with SEQ ID NO 3, said nucleic acid molecule comprising a sequence encoding a bovine Leu-13 protein of about 14 Kda, the expression of said protein being induced by interferon and elevated during bovine pregnancy when compared to levels in non-pregnant bovine animals.
An isolated nucleic acid molecule comprising a sequence selected from the group consisting of:
- a) SEQ ID NO 3;
- b) a sequence which specifically hybridizes with SEQ ID NO 3;
- and
- c) a sequence encoding a polypeptide of SEQ ID NO 4.
An oligonucleotide between about 10 and about 200 nucleotides in length, which specifically hybridizes with a protein translation initiation site in a nucleotide sequence encoding amino acids of SEQ ID NO 4.
An isolated bovine Leu-13 protein, about 14 kDa encoded by SEQ ID NO:
- 4, expression of said encoded protein being induced by interferon and elevated during bovine pregnancy.
A method for detecting pregnancy in a bovine test animal comprising:
- a) obtaining a plurality of biological samples from a test animal and from a non-pregnant animal;
- b) contacting said samples with primers which specifically amplify bovine 1-8U mRNA;
- c) performing polymerase chain reaction on said samples;
- d) detecting amplified bovine 1-8U nucleic acids, an elevation of said 1-8U nucleic acid level obtained from said test animal, relative to that obtained from said non-pregnant animal being indicative of pregnancy in said test animal.
A method for detecting pregnancy in a bovine test animal comprising:
- a) obtaining a plurality of biological samples from a test animal and from a non-pregnant animal;
- b) contacting said samples with primers which specifically amplify bovine Leu-13 mRNA;
- c) performing polymerase chain reaction on said samples;
- d) detecting amplified bovine Leu-13 nucleic acids, an elevation of said Leu-13 nucleic acid level obtained from said test animal, relative to that obtained from said non-pregnant animal being indicative of pregnancy in said test animal.
A method for detecting pregnancy in a bovine test animal comprising:
- a) obtaining a plurality of biological samples from a test animal and from a non-pregnant animal;
- b) contacting said samples with a detectably labeled antibody immunospecific for bovine 1-8U protein;
- d) detecting bovine 1-8U protein, an elevation of said 1-8U protein level obtained from said test animal, relative to that obtained from said non-pregnant animal being indicative of pregnancy in said test animal.
A method for detecting pregnancy in a bovine test animal comprising:
- a) obtaining a plurality of biological samples from a test animal and from a non-pregnant animal;
- b) contacting said samples with a detectably labeled antibody immunospecific for bovine Leu-13 protein;
- d) detecting bovine Leu-13 protein, an elevation of said Leu-13 protein level obtained from said test animal, relative to that obtained from said non-pregnant animal being indicative of pregnancy in said test animal.
Dependencies are inferred from the claim text.