Identification and use of genes encoding amatoxin and phallotoxin
The present invention relates to compositions and methods comprising genes and peptides associated with cyclic peptide toxins and toxin production in mushrooms. In particular, the present invention relates to using genes and proteins from Amanita species encoding Amanita peptides, specifically relating to amatoxins and phallotoxins. In a preferred embodiment, the present invention also relates to methods for detecting Amanita peptide toxin genes for identifying Amanita peptide-producing mushrooms and for diagnosing suspected cases of mushroom poisoning. Further, the present inventions relate to providing kits for diagnosing and monitoring suspected cases of mushroom poisoning in patients.
1. A nucleic acid consisting essentially of one of the sequences set forth in SEQ ID NOs: 55, 56, or 79.
2. A composition comprising at least one isolated nucleic acid consisting of SEQ ID NO: 55, 56, or 79.
3. A method of identifying a toxin-producing mushroom, comprising,
a) providing,
(i) a sample,
(ii) a set of at least two polymerase chain reaction primers with sequences selected from SEQ ID NOs: 1-4, wherein said primers are capable of amplifying an amanitin or phallacidin nucleic acid, and
(iii) a DNA polymerase,
b) mixing said sample with said set of polymerase chain reaction primers,
c) completing a polymerase chain reaction under conditions capable of amplifying an amanitin or phallacidin nucleic acid, and
d) detecting the presence or absence of an amplified amanitin or phallacidin nucleic acid.
4. The method of claim 3 , wherein said sample is selected from the group consisting of a raw sample, a cooked sample, and a digested sample.
5. The method of claim 3 , wherein said sample comprises a mushroom sample.
6. The method of claim 3 , wherein said sample is obtained from a subject.
7. A diagnostic kit for identifying a poisonous mushroom comprising one or more nucleic acids consisting essentially of SEQ ID NO: 55, 56, or 79 and instructions for identifying an amanitin or phallacidin nucleic acid.
8. The kit of claim 7 , wherein said kit further comprises an amanitin or phallacidin nucleic acid consisting of SEQ ID NO: 57, 76, 77, or 81, wherein said nucleic acid is a positive control that can be amplified by a DNA polymerase and polymerase chain reaction primers, a label or a colorimetric reaction product, and instructions for detecting the presence or absence of an amplified nucleic acid.
9. A labeled nucleic acid consisting of any of the sequences set forth in SEQ ID NOs: 57, 76, 77, and 81, and a label.