Direct quantification of ribosome inactivating protein
AssigneeThe United States of America as represented by the Secretary of the Army
InventorRussell M. Dorsey
Life of this patent
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Abstract
A method for detecting ribosome inactivating protein (RIP) activity in a sample is provided that involves contacting a sample that contains an RIP with an inventive substrate that is depurinated to form product. The product is hybridized to a template and cleaved by an apurinic/apyrimidinic endonuclease such that releasing the blocked 3′ end only in a sample that contains RIP activity. The 5′ end of the product serves as a primer for extension by a polymerase reaction. The newly synthesized strand complementary to the template is detected by RT-PCR processes. A kit is provided suitable for field or laboratory use.
Claims16
A process for detecting activity of a ribosome inactivating protein, comprising:
- providing an oligonucleotide substrate consisting of SEQ ID NO:
- 2 with a GAGA loop;
- contacting said substrate with a sample containing a ribosome inactivating protein to produce a product;
- hybridizing said product to a template;
- cleaving said product with an endonuclease;
- extending said product by a polymerization reaction to form an oligonucleotide strand complementary to said template;
- and
- detecting said oligonucleotide strand complementary to said template.
A process for detecting ribosome inactivating protein in a sample, comprising:
- providing an oligonucleotide substrate of SEQ ID NO:
- 1;
- contacting said substrate with a sample containing a ribosome inactivating protein to produce a product;
- hybridizing said product to a template of SEQ ID NO:
- 6;
- cleaving said product with an endonuclease;
- extending said product by a polymerization reaction to form an oligonucleotide strand complementary to said template;
- and
- detecting said oligonucleotide strand complementary to said template by RT-PCR.
Dependencies are inferred from the claim text.