Polynucleotides encoding coccidian parasite casein kinase I, a chemotherapeutic target for antiprotozoal agents
View Patent ↗Isolated nucleic acid molecules encoding coccidian casein kinase I, CKI, enzymes from the species Eimeria tenella and Toxoplasma gondii are disclosed. The isolation of these coccidian CKI cDNA sequences results in the disclosure of purified forms of E. tenella and T. gondii CKI proteins, recombinant vectors and recombinant hosts which express coccidian CKI.
1. A purified nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of:
(a) a nucleotide sequence which encodes the amino acid sequence of SEQ ID NO:2; and
(b) a nucleotide sequence which hybridizes under conditions of high stringency to the complement of a second nucleic acid molecule which encodes the amino acid sequence of SEQ ID NO:2, wherein high stringency conditions are 0.1×SSC at about 68° C.,
wherein the nucleotide sequence is a coccidian sequence and encodes an amino acid sequence that has at least 90% identity to the amino acid sequence of SEQ ID NO:2 and has casein kinase I (CKI) activity.
2. An expression vector for expressing a coccidian CKI protein in a recombinant host cell wherein said expression vector comprises the nucleic acid molecule of claim 1 .
3. An isolated recombinant host cell which expresses a coccidian CKI protein wherein said host cell contains the expression vector of claim 2 .
4. A process of expressing a coccidian CKI protein in a recombinant host cell, comprising:
(a) transfecting the expression vector of claim 2 in a suitable host cell; and,
(b) culturing the isolated host cell of step (a) under conditions which allow expression of said coccidian CKI protein from said expression vector.
5. An isolated nucleic acid molecule encoding a coccidian CKI protein, wherein said nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO:1.
6. An expression vector for expressing a coccidian protein in a recombinant cell wherein said expression vector comprises the nucleic acid molecule of claim 5 .
7. An isolated recombinant host cell which expresses a coccidian CKI protein wherein said host cell contains the expression vector of claim 6 .
8. A process of expressing a coccidian CKI protein in an isolated recombinant host cell, comprising:
(a) transfecting the expression vector of claim 6 into a suitable isolated host cell; and
(b) culturing the host cell of step (a) under conditions which allow expression of said coccidian CKI protein from said expression vector.
9. The purified nucleic acid molecule of claim 1 , wherein said nucleic acid molecule comprises a nucleotide sequence which encodes the amino acid sequence of SEQ ID NO:2.